The present invention relates to the compounds that modulate the activity of the endothelin family of peptides. In particular, the invention relates to the use of sulfonamides and sulfonamide pro-drugs as endothelin agonists and antagonists.
The vascular endothelium releases a variety of vasoactive substances, including the endothelium-derived vasoconstrictor peptide, endothelin (ET) (see, e, Vanhoutte et al. (1986) Annual Rev. Physiol. 48: 307-320; Furchgott and Zawadski (1980) Nature 288: 373-376). Endothelin, which was originally identified in the culture supernatant of porcine aortic endothelial cells (see, Yanagisawa et al. (1988) Nature 332: 411-415), is a potent twenty-one amino acid peptide vasoconstrictor. It is the most potent vasopressor known and is produced by numerous cell types, including the cells of the endothelium, trachea, kidney and brain. Endothelin is synthesized as a two hundred and three amino acid precursor preproendothelin that contains a signal sequence which is cleaved by an endogenous protease to produce a thirty-eight (human) or thirty-nine (porcine) amino acid peptide. This intermediate, referred to as big endothelin, is processed in vivo to the mature biologically active form by a putative endothelin-converting enzyme (ECE) that appears to be a metal-dependent neutral protease (see, e.g., Kashiwabara et al. (1989) FEBS Lttrs. 247: 337-340). Cleavage is required for induction of physiological responses (see, e.g., von Geldern et al. (1991) Peptide Res. 4: 32-35). In porcine aortic endothelial cells, the thirty-nine amino acid intermediate, big endothelin, is hydrolyzed at the Trp21-Val22 bond to generate endothelin-1 and a C-terminal fragment. A similar cleavage occurs in human cells from a thirty-eight amino acid intermediate. Three distinct endothelin isopeptides, endothelin-1, endothelin-2 and endothelin-3, that exhibit potent vasoconstrictor activity have been identified.
The family of three isopeptides endothelin-1, endothelin-2 and endothelin-3 are encoded by a family of three genes (see, Inoue et al. (1989) Proc. Natl. Acad. Sci. USA 86: 2863-2867; see, also Saida et al. (1989) J. Biol. Chem. 264: 14613-14616). The nucleotide sequences of the three human genes are highly conserved within the region encoding the mature 21 amino acid peptides and the C-terminal portions of the peptides are identical. Endothelin-2 is (Trp6, Leu7) endothelin-1 and endothelin-3 is (Thr2,Phe4,Thr5,Tyr6,Lys7,Tyr14) endothelin-1. These peptides are, thus, highly conserved at the C-terminal ends.
Release of endothelins from cultured endothelial cells is modulated by a variety of chemical and physical stimuli and appears to be regulated at the level of transcription and/or translation. Expression of the gene encoding endothelin-1 is increased by chemical stimuli, including adrenaline, thrombin and Ca2+ ionophore. The production and release of endothelin from the endothelium is stimulated by angiotensin II, vasopressin, endotoxin, cyclosporine and other factors (see, Brooks et al. (1991) Eur. J. Pharm. 194:115-117), and is inhibited by nitric oxide. Endothelial cells appear to secrete short-lived endothelium-derived relaxing factors (EDRF), including nitric oxide or a related substance (Palmer et al. (1987) Nature 327: 524-526), when stimulated by vasoactive agents, such as acetylcholine and bradykinin. Endothelin-induced vasoconstriction is also attenuated by atrial natriuretic peptide (ANP).
The endothelin peptides exhibit numerous biological activities in vitro and, in vivo. Endothelin provokes a strong and sustained vasoconstriction in vivo in rats and in isolated vascular smooth muscle preparations; it also provokes the release of eicosanoids and endothelium-derived relaxing factor (EDRF) from perfused vascular beds. Intravenous administration of endothelin-1 and in vitro addition to vascular and other smooth muscle tissues produce long-lasting pressor effects and contraction, respectively (see, em, Bolger et al. (1991) Can. J. Physiol. Pharmacol. 69: 406-413). In isolated vascular strips, for example, endothelin-1 is a potent (EC50=4xc3x9710xe2x88x9210 M), slow acting, but persistent, contractile agent. In vivo, a single dose elevates blood pressure in about twenty to thirty minutes. Endothelin-induced vasoconstriction is not affected by antagonists to known neurotransmitters or hormonal factors, but is abolished by calcium channel antagonists. The effect of calcium channel antagonists, however, is most likely the result of inhibition of calcium influx, since calcium influx appears to be required for the long-lasting contractile response to endothelin.
Endothelin also mediates renin release, stimulates ANP release and induces a positive inotropic action in guinea pig atria. In the lung, endothelin-1 acts as a potent bronchoconstrictor (Maggi et al. (1 989) Eur. J. Pharmacol. 160: 179-182). Endothelin increases renal vascular resistance, decreases renal blood flow, and decreases glomerular filtrate rate. It is a potent mitogen for glomerular mesangial cells and invokes the phosphoinoside cascade in such cells (Simonson et al. (1990) J. Clin. Invest. 85: 790-797).
There are specific high affinity binding sites (dissociation constants in the range of 2-6xc3x9710xe2x88x9210 M) for the endothelins in the vascular system and in other tissues, including the intestine, heart, lungs, kidneys, spleen, adrenal glands and brain. Binding is not inhibited by catecholamines, vasoactive peptides, neurotoxins or calcium channel antagonists. Endothelin binds and interacts with receptor sites that are distinct from other autonomic receptors and voltage dependent calcium channels. Competitive binding studies indicate that there are multiple classes of receptors with different affinities for the endothelin isopeptides. The sarafotoxins, a group of peptide toxins from the venom of the snake Atractaspis eingadensis that cause severe coronary vasospasm in snake bite victims, have structural and functional homology to endothelin-1 and bind competitively to the same cardiac membrane receptors (Kloog et al. (1989) Trends Pharmacol. Sci. 10: 212-214).
Two distinct endothelin receptors, designated ETA and ETB, have been identified and DNA clones encoding each receptor have been isolated (Arai et al. (1990) Nature 348: 730-732; Sakurai et al. (1990) Nature 348: 732-735). Based on the amino acid sequences of the proteins encoded by the cloned DNA, it appears that each receptor contains seven membrane spanning domains and exhibits structural similarity to G-protein-coupled membrane proteins. Messenger RNA encoding both receptors has been detected in a variety of tissues, including heart, lung, kidney and brain. The distribution of receptor subtypes is tissue specific (Martin et al. (1989) Biochem. Biophys. Res. Commun. 162: 130-137). ETA receptors appear to be selective for endothelin-1 and are predominant in cardiovascular tissues. ETB receptors are predominant in noncardiovascular tissues, including the central nervous system and kidney, and interact with the three endothelin isopeptides (Sakurai et al. (1990) Nature 348: 732-734). In addition, ETA receptors occur on vascular smooth muscle, are linked to vasoconstriction and have been associated with cardiovascular, renal and central nervous system diseases; whereas ETB receptors are located on the vascular endothelium, linked to vasodilation (Takayanagi et al. (1991) FEBS Lttrs. 282: 103-106) and have been associated with bronchoconstrictive disorders.
By virtue of the distribution of receptor types and the differential affinity of each isopeptide for each receptor type, the activity of the endothelin isopeptides varies in different tissues. For example, endothelin-1 inhibits 125I-labelled endothelin-1 binding in cardiovascular tissues forty to seven hundred times more potently than endothelin-3. 125I-labelled endothelin-1 binding in non-cardiovascular tissues, such as kidney, adrenal gland, and cerebellum, is inhibited to the same extent by endothelin-1 and endothelin-3, which indicates that ETA receptors predominate in cardiovascular tissues and ETB receptors predominate in non-cardiovascular tissues.
Endothelin plasma levels are elevated in certain disease states (see, e.q., International PCT Application WO 94/27979, and U.S. Pat. No. 5,382,569, which disclosures are herein incorporated in their entirety by reference). Endothelin-1 plasma levels in healthy individuals, as measured by radioimmunoassay (RIA), are about 0.26-5 pg/ml. Blood levels of endothelin-1 and its precursor, big endothelin, are elevated in shock, myocardial infarction, vasospastic angina, kidney failure and a variety of connective tissue disorders. In patients undergoing hemodialysis or kidney transplantation or suffering from cardiogenic shock, myocardial infarction or pulmonary hypertension levels are as high as 35 pg/ml have been observed (see, Stewart et al. (1991) Annals Internal Med. 114: 464-469). Because endothelin is likely to be a local, rather than a systemic, regulating factor, it is probable that the levels of endothelin at the endothelium/smooth muscle interface are much higher than circulating levels.
Elevated levels of endothelin have also been measured in patients suffering from ischemic heart disease (Yasuda et al. (1990) Amer. Heart J. 119:801-806, Ray et al. (1992) Br. Heart J. 67:383-386). Circulating and tissue endothelin immunoreactivity is increased more than twofold in patients with advanced atherosclerosis (Lerman et al. (1991) New Engl. J. Med. 325:997-1001). Increased endothelin immunoreactivity has also been associated with Buerger""s disease (Kanno et al. (1990) J. Amer. Med. Assoc. 264:2868) and Raynaud""s phenomenon (Zamora et al. (1990) Lancet 336 1144-1147). Increased circulating endothelin levels were observed in patients who underwent percutaneous transluminal coronary angioplasty (PTCA) (Tahara et al. (1991) Metab. Clin. Exp. 40:1235-1237; Sanjay et al. (1991) Circulation 84(Suppl. 4):726), and in individuals (Miyauchi et al. (1992) Jpn. J. Pharmacol.58:279P; Stewart et al. (1991) Ann.Internal Medicine 114:464-469) with pulmonary hypertension. Thus, there is clinical human data supporting the correlation between increased endothelin levels and numerous disease states.
Endothelin Agonists and Antagonists
Because endothelin is associated with certain disease states and is implicated in numerous physiological effects, compounds that can interfere with or potentiate endothelin-associated activities, such as endothelin-receptor interaction and vasoconstrictor activity, are of interest. Compounds that exhibit endothelin antagonistic activity have been identified. For example, a fermentation product of Streptomyces misakiensis, designated BE-18257B, has been identified as an ETA receptor antagonist. BE-18257B is a cyclic pen tapeptide, cyclo(D-Glu-L-Ala-allo-D-lle-L-Leu-D-Trp), which inhibits 125I-labelled endothelin-1 binding in cardiovascular tissues in a concentration-dependent manner (IC50 1.4 xcexcM in aortic smooth muscle, 0.8 xcexcM in ventricle membranes and 0.5 xcexcM in cultured aortic smooth muscle cells), but fails to inhibit binding to receptors in tissues in which ETB receptors predominate at concentrations up to 100 xcexcM. Cyclic pentapeptides related to BE-18257B, such as cyclo(D-Asp-Pro-D-Val-Leu-D-Trp) (BQ-123), have been synthesized and shown to exhibit activity as ETA receptor antagonists (see, U.S. Pat. No. 5,114,918 to Ishikawa et al.; see, also, EP A1 0 436 189 to BANYU PHARMACEUTICAL CO., LTD (Oct. 7, 1991)). Studies that measure the inhibition by these cyclic peptides of endothelin-1 binding to endothelin-specific receptors indicate that these cyclic peptides bind preferentially to ETA receptors. Other pepidoe and non-peptide ETA antagonists have been identified (see, e.g., U.S. Pat. Nos. 5,352,800, 5,334,598, 5,352,659, 5,248,807, 5,240,910, 5,198,548, 5,187,195, 5,082,838). These include other cyclic peptides, acyltripeptides, hexapeptide analogs, certain anthraquinone derivatives, indanecarboxylic acids, certain N-pyrimidinylbenzenesulfonamides, certain benzenesulfonamides, and certain naphthalenesulfonamides (Nakajima et al. (1991) J. Antibiot. 44:1348-1356; Miyata et al. (1992) J. Antibiot. 45:74-8; Ishikawa et al. (1992) J.Med. Chem. 35:2139-2142; U.S. Pat. No. 5,114,918 to Ishikawa et al.; EP A1 0 569 193; EP A1 0 558 258; EP A1 0 436 189 to BANYU PHARMACEUTICAL CO., LTD (Oct. 7, 1991); Canadian Patent Application 2,067,288; Canadian Patent Application 2,071,193; U.S. Pat. Nos. 5,208,243; 5,270,313; Cody et al. (1993) Med. Chem. Res. 3:154-162; Miyata et al. (1992) J. Antibiot 45:1041-1046; Miyata et al. (1992) J. Antibiot 45:1029-1040, Fujimoto et al. (1992) FEBS Lett. 305:41-44; Oshashi et al. (1002) J. Antibiot 45:1684-1685; EP A1 0 496 452; Clozel et al. (1993) Nature 365:759-761; International Patent Application WO 93/08799; Nishikibe et al. (1993) Life Sci. 52:717-724; and Benigni et al. (1993) Kidney Int. 44:440-444). In general, the identified compounds have activities in in vitro assays as ETA antagonists at concentrations on the order of about 50-100 xcexcM or less. A number of such compounds have also been shown to possess activity in In vivo animal models. Very few, if any, selective ETB antagonists have been identified.
Endothelin Antagonists and Agonists as Therapeutic Agents
It has been recognized that compounds that exhibit activity at IC50 or EC50 concentrations on the order of 10xe2x88x924 or lower in standard in vitro assays that assess endothelin antagonist or agonist activity have pharmacological utility (see, e., U.S. Pat. Nos. 5,352,800, 5,334,598, 5,352,659, 5,248,807, 5,240,910, 5,198,548, 5,187,195, 5,082,838). By virtue of this activity, such compounds are considered to be useful for the treatment of hypertension such as peripheral circulatory failure, heart disease such as angina pectoris, cardiomyopathy, arteriosclerosis, myocardial infarction, pulmonary hypertension, vasospasm, vascular restenosis, Raynaud""s disease, cerebral stroke such as cerebral arterial spasm, cerebral ischemia, late phase cerebral spasm after subarachnoid hemorrhage, asthma, bronchoconstriction, renal failure, particularly post-ischemic renal failure, cyclosporine nephrotoxicity such as acute renal failure, colitis, as well as other inflammatory diseases, endotoxic shock caused by or associated with endothelin, and other diseases in which endothelin has been implicated.
Thus, in view of the numerous physiological effects of endothelin and its association with certain diseases, endothelin is believed to play a critical role in these pathophysiological conditions (see, e.g., Saito et al. (1990) Hypertension 15: 734-738; Tomita et al. (1989) N. Engl. J. Med. 321: 1127; Kurihara et al. (1989) J. Cardiovasc. Pharmacol. 13(Suppl. 5): S13-S17; Doherty (1992) J. Med. Chem. 35: 1493-1508; Morel et al. (1989) Eur. J. Pharmacol. 167: 427-428). More detailed knowledge of the function and structure of the endothelin peptide family should provide insight in the progression and treatment of such conditions.
To aid in gaining further understanding of and to develop treatments for endothelin-mediated or related disorders, there is a need to identify compounds that modulate or alter endothelin activity. Identification of compounds that modulate endothelin activity, such as compounds that act as specific antagonists or agonists, may not only aid in elucidating the function of endothelin, but may yield in therapeutically useful compounds. In particular, compounds that specifically interfere with the interaction of endothelin peptides with ETA, ETB or other receptors should be useful in identifying essential characteristics of endothelin peptides, should aid in the design of therapeutic agents, and may be useful as disease specific therapeutic agents.
Therefore, it is an object herein to provide compounds that have the ability to modulate the biological activity of one or more of the endothelin isopeptides. It is another object to provide compounds that have use as specific endothelin antagonists. It is of particular interest herein to provide compounds that selectively interact with ETB receptors. It is also an object to use compounds that specifically interact with or inhibit the interaction of endothelin peptides with ETA or ETB receptors. Such compounds should be useful as therapeutic agents for the treatment of endothelin-mediated diseases and disorders and also for the identification of endothelin receptor subtypes.
Sulfonamides and methods for modulating the interation of an endothelin peptide with ETA and/or ETB receptors are provided. In particular, sulfonamides and methods for inhibiting the binding of an endothelin peptide to ETA or ETB receptors are provided. Sulfonamides that act as endothelin peptide agonists with respect to ETA or ETB receptors are also provided. Among the compounds provided herein are those that are particularly active as ETB antagonists.
The methods are effected by contacting endothelin receptors with one or more sulfonamides prior to, simultaneously with, or subsequent to contacting the receptors with an endothelin peptide. The sulfonamides are substituted or unsubstituted monocyclic or polycyclic aromatic or heteroaromatic sulfonamides, such as benzene sulfonamides, particularly biphenyl sulfonamides, napthalene sulfonamides, and fused tricyclic ring sulfonamides.
The sulfonamides have formula I: 
in which
Ar1 is a substituted or unsubstituted aryl group with one or more substituents, including an alkyl group, an aryl group, a substituted aryl group, a nitro group, an amino group or a halide or is an alkyl group. In particular, Ar1 is alkyl or is a five or six membered substituted or unsubstituted aromatic or heteroaromatic rings, including, 3- or 5-isoxazolyl, thiazolyl such as 2-thiazolyl, pyrimidinyl such as 2-pyrimidinyl, pyridazinyl, and unsubstituted or substituted benzene groups, including aryloxy substituted benzene groups or is a bicyclic or tricyclic ring.
Ar2 is any group such that the resulting sulfonamide inhibits binding by 50%, compared to binding in the absence of the sulfonamide, of an endothelin peptide to an endothelin receptor at a concentration of the sulfonamide of less than about 100 xcexcM, except that Ar2 is not phenyl, 2-biphenyl or naphthyl unless R1 is a halide or higher alkyl or unless it is a biphenyl, other than a 2-biphenyl, and the substituent at the 2and/or 6 position(s) on the phenyl linked to the sulfonamide is hydrogen.
Ar1 is, in certain embodiments, selected from groups such as: 
and R is selected from H, NH2, halide, pseudohalide, alkyl alkylcarbonyl, formyl, an aromatic or heteroaromatic group, alkoxyalkyl, alkylamino, alkylthio, arylcarbonyl, aryloxy, arylamino, arylthio, haloalkyl, haloaryl, carbonyl, in which the aryl and alkyl portions, are unsubstituted or substituted with any of the preceding groups, and unsubstituted or substituted with any of the preceding groups, and straight or branched chains of from about 1 up to about 10-12 carbons, preferably, 1 to about 5 or 6 carbons. R is preferably H, NH2, halide, CH3, CH3O or another aromatic group.
In the embodiments described in detail herein, Ar1 is generally an isoxazole and the compounds are represented by the, Formulae II: 
in which R1 and R2 are either (i), (ii) or (iii) as follows:
(i) R1 and R2 are each independently selected from H, NH2, NO2, halide, pseudohalide, alkyl, alkenyl, alkynyl, aryl, arylalkyl, heteroaryl, alkoxy, alkylamino, alkylthio, alkyloxy, haloalkyl, alkylsulfinyl, alkylsulfonyl, aryloxy, arylamino, arylthio, arylsulfinyl, arylsulfonyl, haloalkyl, haloaryl, alkoxycarbonyl, alkylcarbonyl, aminocarbonyl, arylcarbonyl, formyl, substituted or unsubstituted amido, substituted or unsubstituted ureido, in which the alkyl, alkenyl and alkynyl portions contain from 1 up to about 14 carbon atoms and are either straight or branched chains or cyclic, and the aryl portions contain from about 4 to about 16 carbons, except that R2 is not halide or pseudohalide; or,
(ii) R1 and R2 together form xe2x80x94(CH2)n, where n is 3 to 6; or,
(iii) R1 and R2 together form 1,3-butadienyl, and with the proviso that Ar2 is not 2-biphenyl, naphthyl or phenyl, other than certain 3-biphenyls or 4-biphenyls, unless R1 is a halide or higher (C8-C15, preferably C9-C13) alkyl. R1 more preferably bromide or chloride, methyl or ethyl or (C9-C13)alkyl. In the most active compounds provided herein, as evidenced by in vitro binding assays, R1 is bromide or chloride or, in instances in which ETB selectivity is desired, C9-C13 alkyl.
In other embodiments herein, Ar1 is pyridazinyl, particularly N-3-pyridazinyl sulfonamides, in which the pyridazinyl group is unsubstituted or substituted with one or more substituents selected from R.
In the embodiments described in detail herein, Ar2 is naphthyl, phenyl, including biphenyl or a group that is a derivative of or analog, as described below, of a biphenyl group, and Ar1 is preferably N-(5-isoxazolyl) or N-(3-isoxazolyl) or N-pyridazinyl, preferably N-3-pyridazinyl.
In the preferred compounds herein, R2 is preferably, selected from among alkyl, lower alkenyl, lower alkynyl, lower haloalkyl, halide or H; and R1 is halide, lower alkyl, or higher (C8-C15, preferably C9-C13) alkyl, except: (a) when Ar2 is phenyl and R4, R5, R6 and R7 are hydrogen, R3 is not, NO2, NH2 or lower alkyl, unless R1 is higher alkyl; and (b) if Ar2 is naphthyl or if Ar2 is phenyl, or if Ar2 is 2-biphenyl or if Ar2 is 3-biphenyl in which R4 and R5 are other than hydrogen, then R1 is halide or higher alkyl (C8H17xe2x80x94C15H29), preferably C9H17xe2x80x94C13H27).
In particular embodiments disclosed herein, Ar2 is a group of formulae III: 
in which:
n is 0 to 10, preferably 0 to 3, more preferably 0 or 1, most preferably 0;
R3, R4, R5, R6, and R7 are selected from (i), (ii), (iii), (iv) or (v) with the proviso that: (a) when Ar2 is phenyl and R4, R5, R6 and R7 are hydrogen, R3is not NO2, NH2 or lower alkyl, unless R1 is higher alkyl; (b) if Ar2 is naphthyl or if Ar2 is phenyl, or if Ar2 is 2-biphenyl or if Ar2 is 3-biphenyl in which R4 and R5 are other than hydrogen, then R1 is halide or higher (C8-C15, preferably C9-C13) alkyl:
(i) R3, R4, R5, R6, and R7 are each selected independently from among H, NHOH, NH2, NO2, N3, halide, pseudohalide, alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkoxy, alkylamino, alkylthio, alkoxyalkyl, alkylsulfinyl, alkylsulfonyl, aryloxy, arylamino, arylthio, arylsulfinyl, arylsulfonyl, haloalkyl, haloaryl, alkoxycarbonyl, alkylcarbonyl, arylcarbonyl, formyl, substituted or unsubstituted amido, substituted or unsubstituted ureido, where the alkyl, alkenyl, alkynyl portions are straight or branched chains of from about 1 up to about 10 carbons, preferably, 1 to about 5 or 6 carbons and the aryl portions contain from 3 up to about 10 carbons, preferably 6 carbons; or, alternatively,
(ii) R4 and R7 together are substituted or unsubstituted 1,3-butadienyl, 4-dimethylamino-1,3-butadienyl, 1-chloro-1,3-butadienyl, 1-aza-1,3-butadienyl or 2-aza-1,3-butadienyl groups; and R3, R5 and R6 are as defined in (i) above; or alternatively,
(iii) R7 and R3 together are substituted or unsubstituted 1,3-butadienyl, 4-dimethylamino-1,3-butadienyl, 1-chloro-1,3-butadienyl, 1-aza-1,3-butadienyl or 2-aza-1,3-butadienyl groups; and n, X, R4, R5 and R6 or as defined in (i) above; or
(iv) R3, R5, and R7are H are as defined in (i); and R4 and R6 are each independently selected from alkyl, alkoxy, halide aminoalkyl, dialkylaminoalkyl, in which the alkyl and alkoxy groups contain from 1 to 10, preferably 1 to 6 carbons, and are straight or branched chains; or
(v) any two of R3, R4, R5, R6, and R7, which are each selected as in (i) form fused carbocyclic or heterocyclic rings.
Thus, in the compounds provided herein Ar2 is selected from among phenyl, biphenyl, naphthyl, and bicyclic and tricyclic fused carbocyclic and heterocyclic rings, and other such groups.
Selected isoxazolyl-benzenesulfonamides and isoxazolylnaphthalenesulfonamides in which the isoxazole is other than a 4-halo-isoxazole are also provided. Such selected compounds, including N-isoxazolylbenzenesulfonamides and N-isoxazolylnaphthalenesulfonamides in which the substituent at the 4 position on the isoxazolyl group is higher alkyl, such as C9H19 to C13H27 are also provided. These compounds have enhanced ETB affinity compared to corresponding compounds in which the substituent at the 4 position is lower alkyl or other groups, such as pseudohalide, halide, alkylaryl, aryl, lower alkyl, carboxamide, alkoxy, and others.
In embodiments in which Ar2 is biphenyl, one of R3, R4, R5, R6 and R7 is phenyl, which is substituted or is unsubstituted with substituents such as Z1-Z5 in which each Z is independently selected from hydrogen, halide, pseudohalide, alkyl, alkoxy, alkenyl, alkynyl, aryl, heterocycle, aralkyl, aralkoxy, cycloalkyl, cycloalkenyl, cycloalkynyl, OH, CN, C(O)R16, CO2R16, SH, S(O)nR16 in which n is 0-2, NHOH, NR12R 16, NO2, N3, OR16, R12NCOR16 and CONR12R16; and R12 and R16 are each independently selected from hydrogen, alkyl, alkenyl, alkynyl, aryl, alkylaryl, heterocycle, aralkyl, aralkoxy, cycloalkyl, cycloalkenyl or cycloalkynyl; and
the others of R3, R4, R5, R6 and R7 are each independently selected from among H, NHOH, NH2, NO2, N3, halide, pseudohalide, alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkoxy, alkylamino, alkylthio, alkoxyalkyl, alkylsulfinyl, alkylsulfonyl, aryloxy; arylamino, arylthio, arylsulfinyl, arylsulfonyl, haloalkyl, haloaryl, alkoxycarbonyl, alkylcarbonyl, arylcarbonyl, formyl, substituted or unsubstituted amido, substituted or unsubstituted ureido, where the alkyl, alkenyl, alkynyl portions are straight or branched chains of from about 1 up to about 10 carbons, preferably, 1 to about 5 or 6 carbons and the aryl portions contain from 3 up to about 10 carbons, preferably 6 carbons; or, alternatively, with the proviso that: if Ar2 is 2-biphenyl or if Ar2 is 3-biphenyl in which R4 and R5 are other than hydrogen, then R1 is halide or higher (C8-C15, preferably C9-C13) alkyl. In other embodiments, if Ar2 is 4-biphenyl and R4 and/or R5 are other hydrogen, then R1 is halide or higher (C8-C15, preferably C9-C13) alkyl.
Z is preferably selected from:
(i) hydrogen, OH, NH2, NO2, alkyl, haloalkyl, halide, pseudohalide, alkoxy, alkoxyalkyl, NR11, in which R11 is selected from among H, a straight or branched carbon chain, preferably containing 1 to 6, more preferably 1 to 3, carbons, halide, alkoxyalkyl, haloalkyl, S or O, or the like; or
(ii) any two of Z1-Z5 together with the atoms to which each is attached form a carbon ring or heterocycle fused to the phenyl group. It is preferable that at least three or four of Z1-Z5 is hydrogen, and preferably the hydrogens are in the meta positions and ortho positions. Preferred positions for Z1-Z5, when not hydrogen, are the para position and/or ortho position.
In particular, R11 contains up to about 30 carbon atoms, preferably 1 to 10, more preferably 1 to 6 and is selected from hydrogen, alkyl, alkenyl, alkynyl, aryl, alkylaryl, heterocycle, aralkyl, aralkoxy, cycloalkyl, cycloalkenyl, cycloalkynyl, C(O)R15 and S(O)nR15 in which n is 0-2; R15 is hydrogen, alkyl, alkenyl, alkynyl, aryl, alkylaryl, heterocycle, aralkyl, aralkoxy, cycloalkyl, cycloalkenyl, cycloalkynyl; R11 and R15 are unsubstituted or are substituted with one or more substituents each selected independently from Z, which is hydrogen, halide, pseudohalide, alkyl, alkoxy, alkenyl, alkynyl, aryl, heterocycle, aralkyl, aralkoxy, cycloalkyl, cycloalkenyl, cycloalkynyl, OH, CN, C(O)R16, CO2R16, SH, S(O)nR16 in which n is 0-2, NHOH, NR12R16, NO2, N3, OR16, R12NCOR16 and CONR12R16 is hydrogen, alkyl, alkenyl, alkynyl, aryl, alkylaryl, heterocycle, aralkyl, aralkoxy, cycloalkyl, cycloalkenyl or cycloalkynyl; R12, which is selected independently from R11 and Z, is selected from hydrogen, alkyl, alkenyl, alkynyl, aryl, alkylaryl, heterocycle, aralkyl, aralkoxy, cycloalkyl, cycloalkenyl, cycloalkynyl, C(O)R17 and S(O)nR17 in which n is 0-2; and R17 is hydrogen, alkyl, alkenyl, alkynyl, aryl, alkylaryl, heterocycle, aralkyl, aralkoxy, cycloalkyl, cycloalkenyl or cycloalkynyl; each of R11, R12, R15 and R16 may be further substituted with any of hydrogen, halide, pseudohalide, alkyl, alkoxy, alkenyl, alkynyl, aryl, heterocycle, aralkyl, aralkoxy, cycloalkyl, cycloalkenyl, cycloalkynyl, OH and CN.
In preferred embodiments herein, R3 or R6 or R7 is phenyl. In more preferred embodiments, R3 is phenyl, and Z2-Z5 are hydrogen or lower alkyl, preferably C1-3 alkyl, and Z1 is in the ortho, or para position, and is preferably lower alkyl, lower alkoxy, halo-lower alkyl or halide.
Compounds in which Ar2 is a tricyclic caribocycle or heterocycle are also provided. In particular, preferred among these compounds are compounds of formula: 
in which X is S, O, NR14 in which R14 like R11, but selected independently from R11, is aryl, hydrogen, or lower alkyl, preferably, a substituted or unsubstituted aryl, particularly phenyl, preferably unsubstituted or substituted with lower alkyl or halogen hydrogen or lower alkyl, in which the alkyl groups contain from 1-6 carbons, preferably 1-3 carbons, and are straight, branched or cyclic chains.
In the above compounds, the alkyl, alkynyl and alkenyl portions of each listed substituent are straight or branched chains or are cyclic, and preferably have from about 1 up to about 10 carbons; in more preferred embodiments they have from 1-6 carbons, and they can have fewer than 6 carbons. The aryl, carbocyclic, aromatic rings and heterocyclic groups can have from 3 to 16, generally, 3-7, more often 5-7 members in the rings, and may be single or fused rings. The ring size and carbon chain length are selected up to a size such that the resulting molecule binds to retains activity as an endothelin antagonist or agonist, such that the resulting compound inhibits binding by 50%, compared to binding in the absence of the sulfonamide, of an endothelin peptide to an endothelin receptor at a concentration of less than about 100 xcexcM.
Of particular interest herein are compounds that inhibit binding of endothelin to ETB receptors at lower concentrations, preferably at least 2-fold, more preferably 3-fold, and most preferably at least 5- to 10-fold lower than they inhibit binding to ETA receptors.
Preferred compounds are ETB receptor selective or bind to ETB receptors with an IC50 of less than about 1 xcexcM.
Preferred compounds also include compounds that are ETB receptor selective or that competitively inhibit binding of endothelin-1 to ETB receptors at IC50 concentrations of 1 xcexcM (if measured at 4xc2x0 C.; or 5-10 xcexcM, if measured at 24xc2x0 C.). In preferred compounds, generally R2 is selected from among alkyl, lower alkenyl, lower alkynyl, lower haloalkyl, halide or H; and R1 is halide or lower alkyl, and in preferred embodiments, R1 is bromide or chloride.
Of the compounds described herein, those that inhibit or increase an endothelin-mediated activity by about 50% at concentrations of less than about 10 xcexcM are preferred. More preferred are those that inhibit or increase an endothelin-mediated activity by about 50% at concentrations of less than about 1 uM, more preferably less than about 0.1 xcexcM, even more preferably less than about 0.01 xcexcM, and most preferably less than about 0.005 xcexcM. The preferred IC50 concentrations are set forth with reference to the in vitro assays exemplified herein. It is understood that these IC50 concentrations vary from assay to assay. For example, it is noted that, as described below, the IC50 concentration determined in the in vitro assays is a non-linear function of incubation temperature. The preferred values recited herein refer to the assays that are performed at 4xc2x0 C. When the assays are performed at 24xc2x0 C., somewhat higher (see, Table 1) IC50 concentrations are observed. Accordingly, when the assay is performed at 24xc2x0 C., the preferred IC50 concentrations are about 10-fold higher.
Pharmaceutical compositions formulated for administration by an appropriate route and means containing effective concentrations of one or more of the compounds provided herein or pharmaceutically acceptable salts or acids thereof that deliver amounts effective for the treatment of bronchoconstrictive disorders, and other conditions that are in some manner mediated by an endothelin peptide or that involve bronchoconstriction or whose symptoms can be ameliorated by administration of an ETB-specific endothelin antagonist or agonist, are also provided. Particularly preferred compositions are those that deliver amounts effective for the treatment of hypertension or renal failure. The effective amounts and concentrations are effective for ameliorating any of the symptoms of any of the disorders.
Methods for inhibiting binding of an endothelin peptide to an endothelin receptor are provided. These methods are practiced by contacting the receptor with one or more of the compounds provided herein simultaneously, prior to, or subsequent to contacting the receptor with an endothelin peptide.
Methods for treatment of endothelin-mediated disorders, including but not limited to, asthma, conditions that are in some manner mediated by an endothelin peptide that binds to ETB receptors, or for treatment of disorder that involves bronchoconstriction or that are ameliorated by administration of an ETB receptor endothelin antagonist or agonist are provided.
In particular, methods of treating endothelin-mediated disorders by administering effective amounts of the sulfonamides, prodrugs or other suitable derivatives of the sulfonamides are provided. In particular, methods for treating endothelin-mediated disorders, including respiratory diseases and inflammatory diseases, including asthma, bronchioconstriction, and other diseases in which ETB receptor endothelin mediated physiological responses are implicated, by administering effective amounts of one or more of the compounds provided herein in pharmaceutically acceptable carriers are provided. Preferred methods of treatment are methods for treatment of hypertension and renal failure.
More preferred methods of treatment are those in which the compositions contain at least one compound that inhibits the interaction of an endothelin with ETB receptors at an IC50 of less than about 10 xcexcM, and preferably less than about 5 xcexcM, more preferably less than about 1 xcexcM, even more preferably less than 0.1 xcexcM, and most preferably less than 0.05 xcexcM. Other preferred methods are those in which the compositions contain one or more compounds that is (are) ETB selective. Methods in which the compounds are ETB selective are for treatment of disorders, such as asthma, that require bronchodilation.
In practicing the methods, effective amounts of compositions containing therapeutically effective concentrations of the compounds formulated for oral, intravenous, local and topical application for the treatment of hypertension, cardiovascular diseases, cardiac diseases, including myocardial infarction, respiratory diseases, including asthma, inflammatory diseases, ophthalmologic diseases, gastroenteric diseases, renal failure, immunosuppressant-mediated renal vasoconstriction, erythropoietin-mediated vasoconstriction, endotoxin shock, anaphylactic shock, hemorrhagic shock, pulmonary hypertension, and other diseases in which endothelin mediated physiological responses are implicated are administered to an individual exhibiting the symptoms of one or more of these disorders. The amounts are effective to ameliorate or eliminate one or more symptoms of the disorders.
Methods for the identification and isolation of endothelin receptor subtypes, particularly ETB subtypes are also provided. In particular, methods for detecting, distinguishing and isolating endothelin receptors using the disclosed compounds are provided. In particular, methods are provided for detecting, distinguishing and isolating endothelin receptors using the compounds provided herein.
In addition, methods for identifying compounds that are suitable for use in treating particular diseases based on their preferential affinity for a particular endothelin receptor subtype are also provided.
Articles of manufacture containing packaging material, a compound provided herein, which is effective for ameliorating the symptoms of an endothelin-mediated disorder, antagonizing the effects of endothelin or inhibiting binding of an endothelin peptide to an ETB receptor with an IC50 of less than about 10 xcexcM, within the packaging material, and a label that indicates that the compound or salt thereof is used for antagonizing the effects of endothelin, treating an endothelin-mediated disorder, or inhibiting the binding of an endothelin peptide to an ET receptor are provided.
Definitions
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art to which this invention belongs. All patents and publications referred to herein are incorporated by reference.
As used herein, endothelin (ET) peptides include peptides that have substantially the amino acid sequence of endothelin-1, endothelin-2 or endothelin-3 and that act as potent endogenous vasoconstrictor peptides.
As used herein, an endothelin-mediated condition is a condition that is caused by abnormal endothelin activity or one in which compounds that inhibit endothelin activity have therapeutic use. Such diseases include, but are not limited to hypertension, cardiovascular disease, asthma, inflammatory diseases, ophthalmologic disease, menstrual disorders, obstetric conditions, gastroenteric disease, renal failure, pulmonary hypertension, endotoxin shock, anaphylactic shock, or hemorrhagic shock. Endothelin-mediated conditions also include conditions that result from therapy with agents, such as erythropoietin and immunosuppressants, that elevate endothelin levels.
As used herein an effective amount of a compound for treating a particular disease is an amount that is sufficient to ameliorate, or in some manner reduce the symptoms associated with the disease. Such amount may be administered as a single dosage or may be administered according to a regimen, whereby it is effective. The amount may cure the disease but, typically, is administered in order to ameliorate the symptoms of the disease. Typically, repeated administration is required to achieve the desired amelioration of symptoms.
As used herein, an endothelin agonist is a compound that potentiates or exhibits a biological activity associated with or possessed by an endothelin peptide.
As used herein, an endothelin antagonist is a compound, such as a drug or an antibody, that inhibits endothelin-stimulated vasoconstriction and contraction and other endothelin-mediated physiological responses. The antagonist may act by interfering with the interaction of the endothelin with an endothelin-specific receptor or by interfering with the physiological response to or bioactivity of an endothelin isopeptide, such as vasoconstriction. Thus, as used herein, an endothelin antagonist interferes with endothelin-stimulated vasoconstriction or other response or interferes with the interaction of an endothelin with an endothelin-specific receptor, such as ETA receptors, as assessed by assays known to those of skill in the art.
The effectiveness of potential agonists and antagonists can be assessed using methods known to those of skill in the art. For example, endothelin agonist activity can be identified by its ability to stimulate vasoconstriction of isolated rat thoracic aorta or portal vein ring segments (Borges et al. (1989) xe2x80x9cTissue selectivity of endothelinxe2x80x9d Eur. J. Pharmacol. 165: 223-230). Exemplary assays are set forth in the EXAMPLES. As noted above, the preferred IC50 concentration ranges are set forth with reference to assays in which the test compound is incubated with the ET receptor-bearing cells at 4xc2x0 C. Data presented for assays in which the incubation step is performed at the less preferred 24xc2x0 C. are identified. It is understood that for purposes of comparison, these concentrations are somewhat higher than the concentrations determined at 4xc2x0 C.
As used herein, the biological activity or bioactivity of endothelin includes any activity induced, potentiated or influenced by endothelin in vivo. It also includes the ability to bind to particular receptors and to induce a functional response, such as vasoconstriction. It may be assessed by in vivo assays or by in vitro assays, such as those exemplified herein. The relevant activities include, but are not limited to, vasoconstriction, vasorelaxation and bronchodilation. For example, ETB receptors appear to be expressed in vascular endothelial cells and may mediate vasodilation and other such responses; whereas ETA receptors, which are endothelin-1-specific, occur on smooth muscle and are linked to vasoconstriction Any assay known to those of skill in the art to measure or detect such activity may be used to assess such activity (see, e.g., Spokes et al. (1989) J. Cardiovasc. Pharmacol. 13(Suppl. 5):S191-S192; Spinella et al. (1991) Proc. Natl. Acad. Sci. USA 88: 7443-7446; Cardell et al. (1991) Neurochem. Int. 18:571-574); and the Examples herein).
As used herein, the IC50 refers to an amount, concentration or dosage of a particular test compound that achieves a 50% inhibition of a maximal response, such as binding of endothelin to tissue receptors, in an assay that measures such response.
As used herein, EC50 refers to a dosage, concentration or amount of a particular test compound that elicits a dose-dependent response at 50% of maximal expression of a particular response that is induced, provoked or potentiated by the particular test compound.
As used herein a sulfonamide that is ETA selective refers to sulfonamides that exhibit an IC50 that is at least about 10-fold lower with respect to ETA receptors than ETB receptors.
As used herein, a sulfonamide that is ETB selective refers to sulfonamides that exhibit an IC50 that is at least about 10-fold lower with respect to ETB receptors than ETA receptors.
As used herein, pharmaceutically acceptable salts, esters or other derivatives of the compounds include any salts, esters or derivatives that may be readily prepared by those of skill in this art using known methods for such derivatization and that produce compounds that may be administered to animals or humans without substantial toxic effects and that either are pharmaceutically active or are prodrugs. For example, hydroxy groups can be esterified or etherified.
As used herein, treatment means any manner in which the symptoms of a conditions, disorder or disease are ameliorated or otherwise beneficially altered. Treatment also encompasses any pharmaceutical use of the compositions herein, such as use as contraceptive agents.
As used herein, amelioration of the symptoms of a particular disorder by administration of a particular pharmaceutical composition refers to any lessening, whether permanent or temporary, lasting or transient that can be attributed to or associated with administration of the composition.
As used herein, substantially pure means sufficiently homogeneous to appear free of readily detectable impurities as determined by standard methods of analysis, such as thin layer chromatography (TLC), gel electrophoresis and high performance liquid chromatography (HPLC), used by those of skill in the art to assess such purity, or sufficiently pure such that further purification would not detectably alter the physical and chemical properties, such as enzymatic and biological activities, of the substance. Methods for purification of the compounds to produce substantially chemically pure compounds are known to those of skill in the art. A substantially chemically pure compound may, however, be a mixture of stereoisomers. In such instances, further purification might increase the specific activity of the compound.
As used herein, biological activity refers to the in vivo activities of a compound or physiological responses that result upon In vivo administration of a compound, composition or other mixture. Biological activity, thus, encompasses therapeutic effects and pharmaceutical activity of such compounds, compositions and mixtures.
As used herein, a prodrug is a compound that, upon in vivo administration, is metabolized or otherwise converted to the biologically, pharmaceutically or therapeutically active form of the compound. To produce a prodrug, the pharmaceutically active compound is modified such that the active compound will be regenerated by metabolic processes. The prodrug may be designed to alter the metabolic stability or the transport characteristics of a drug, to mask side effects or toxicity, to improve the flavor of a drug or to alter other characteristics or properties of a drug. By virtue of knowledge of pharmacodynamic processes and drug metabolism in vivo, those of skill in this: art, once a pharmaceutically active compound is known, can design prodrugs of the compound (see, e.g., Nogrady (1985) Medicinal Chemistry A Biochemical Approach, Oxford University Press, New York, pages 388-392). For example, succinyl-sulfathiazole is a prodrug of 4-amino-N-(2-thiazolyl)benzenesulfonamide (sulfathiazole) that exhibits altered transport characteristics.
As used herein, xe2x80x9chalogenxe2x80x9d or xe2x80x9chalidexe2x80x9d refers to F, Cl, Br or I.
As used herein, pseudohalides are compounds that behave substantially similar to halides. Such compounds can be used in the same manner and treated in the same manner as halides (X, in which X is a halogen, such as Cl or Br). Pseudohalides include, but are not limited to cyanide, cyanate, thiocyanate, selenocyanate and azide.
As used herein, lower alkyl, lower alkenyl, and lower alkynyl refer to carbon chains having less than about 6 carbons. In preferred embodiments of the compounds provided herein that include alkyl, alkenyl, or alkynyl portions include lower alkyl, lower alkenyl, and lower alkynyl portions.
As used herein aryl refers to cyclic groups containing from 3 to 15 or 16 carbon atoms, preferably from 5 to 10. Aryl groups include, but are not limited to groups, such as phenyl, substituted phenyl, naphthyl, substituted naphthyl, in which the substituent is lower alkyl, halogen, or lower alkoxy. Preferred aryl groups are lower aryl groups that contain less than 7 carbons in the ring structure.
As used herein, the nomenclature alkyl, alkoxy, carbonyl, etc. are used as is generally understood by those of skill in this art. For example, as used herein alkyl refers to saturated carbon chains that contain one or more carbons; the chains may be straight or branched or include cyclic portions or be cyclic. As used herein, alicyclic refers to aryl groups that are cyclic.
As used herein, xe2x80x9chaloalkylxe2x80x9d refers to a lower alkyl radical in which one or more of the hydrogen atoms are replaced by halogen including, but not limited to, chloromethyl, trifluoromethyl, 1-chloro-2-fluoroethyl and the like.
As used herein, xe2x80x9chaloalkoxyxe2x80x9d refers to ROxe2x80x94 in which R is a haloalkyl group.
As used herein, xe2x80x9caminocarbonylxe2x80x9d refers to xe2x80x94C(O)NH2.
As used herein, xe2x80x9calkylaminocarbonylxe2x80x9d refers to xe2x80x94C(O)NHR in which R is hydrogen, alkyl, preferably lower alkyl or aryl, preferably lower aryl.
As used herein xe2x80x9cdialkylaminocarbonylxe2x80x9d as used herein refers to xe2x80x94C(O)NRxe2x80x2R in which Rxe2x80x2 and R are independently selected from alkyl or aryl, preferably lower alkyl or lower aryl; xe2x80x9ccarboxamidexe2x80x9d refers to groups of formula NRxe2x80x2COR.
As used herein, xe2x80x9calkoxycarbonylxe2x80x9d as used herein refers to xe2x80x94C(O)OR in which R is alkyl, preferably lower alkyl or aryl, preferably lower aryl.
As used herein, xe2x80x9calkoxyxe2x80x9d and xe2x80x9cthioalkoxyxe2x80x9d refer to ROxe2x80x94 and RSxe2x80x94, in which R is alkyl, preferably lower alkyl or aryl, preferably lower aryl.
As used herein, xe2x80x9chaloalkoxyxe2x80x9d refers to ROxe2x80x94 in which R is a haloalkyl group.
As used herein, xe2x80x9caminocarbonylxe2x80x9d refers to xe2x80x94C(O)NH2.
As used herein, xe2x80x9calkylaminocarbonylxe2x80x9d refers to xe2x80x94C(O)NHR in which R is alkyl, preferably lower alkyl or aryl, preferably lower aryl.
As used herein, xe2x80x9calkoxycarbonylxe2x80x9d refers to xe2x80x94C(O)OR in which R is alkyl, preferably lower alkyl.
As used herein, cycloalkyl refers to saturated cyclic carbon chains; cycloalkyenyl and cycloalkynyl refer to cyclic carbon chains that include at least one unsaturated triple bond. The cyclic portions of the carbon chains may include one ring or two or more fused rings.
As used herein, heterocycle or heteroaryl refers to ring structures that include at least one carbon atom and one or more atoms, such as N, S and O. The rings may be single rings or two or more fused rings. Heteroaryl is used interchangeably with heterocycle.
As used herein, the abbreviations for any protective groups, amino acids and other compounds, are, unless indicated otherwise, in accord with their common usage, recognized abbreviations, or the IUPAC-IUB Commission on Biochemical Nomenclature (see, (1972) Biochem. 11:942).
A. Compounds for Use in Treating Endothelin-mediated Diseases Isoxazolylsulfonamides in Which Ar2 is Phenyl, Biphenyl and Fused Carbocyclic and Heterocyclic Rings
Compounds and methods for treating endothelin-mediated diseases using the compounds of formula I are provided. Compounds in which Ar2 is selected from phenyl, biphenyl, and aromatic fused rings, including naphthyl, anthracenyl, phenanthryl, indenyl, azulenyl, fluorenyl, and phenazinyl and in which Ar1 is oxazolyl or other heterocycle, such as pyridazinyl, are provided.
In particular, the compounds provided herein have formulae II. When Ar2 is phenyl, biphenyl or naphthyl, the compounds are preferably (4-halo-isoxazolyl)sulfonamides or are (4-higher alkyl-isoxazolyl)sulfonmides, in which the alkyl group contains more than about 8, preferably 9 to 15, more preferably 9 to 13, carbon atoms.
Among the compounds provided herein are those of formulae (III): 
in which n is 0 to 10, preferably 0 to 6, more preferably 0 to 3; R3, R4, R5, R6, and R7 are selected from (i), (ii), (iii), (iv) or (v) with the proviso that: (a) when Ar2 is phenyl, and R4, R5, R6 and R7 are hydrogen, R3is not NO2, NH2 or lower alkyl, particularly CH3, unless R1 is higher alkyl; and (b) when Ar2 is naphthyl, 2-biphenyl or phenyl, other than 3- or 4-biphenyl in which R4 and R5 are hydrogen or fused rings (except for naphthyl), R1 is halide or higher alkyl (C8-C15 preferably C9-C13):
(i) R3, R4, R5, R6, and R7 are each selected independently from among H, NHOH, NH2, NO2, N3, halide, pseudohalide, alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkoxy, alkylamino, alkylthio, alkoxyalkyl, alkylsulfinyl, alkylsulfonyl, aryloxy, arylamino, arylthio, arylsulfinyl, arylsulfonyl, haloalkyl, haloaryl, alkoxycarbonyl, alkylcarbonyl, arylcarbonyl, formyl, substituted or unsubstituted amido, substituted or unsubstituted ureido, where the alkyl, alkenyl, alkynyl portions are straight or branched chains of from about 1 up to about 10 carbons, preferably, 1 to about 5 or 6 carbons and the aryl portions contain from 3 up to about 10 carbons, preferably 6 carbons; or, alternatively,
(ii) R4 and R7 together are substituted or unsubstituted 1,3-butadienyl, 4-dimethylamino-1,3-butadienyl, 1-chloro-1,3-butadienyl, 1-aza-1,3-butadienyl or 2-aza-1,3-butadienyl groups; and R3, R5 and R6 are as defined in (i) above; or alternatively,
(iii) R7 and R3 together are substituted or unsubstituted 1,3-butadienyl, 4-dimethylamino-1,3-butadienyl, 1-chloro-1,3-butadienyl, 1-aza-1,3-butadienyl or 2-aza-1,3-butadienyl groups; and n, X, R4, R5 and R6 are as defined in (i) above; or
(iv) R3, R5, and R7 or H or as defined in (i); and R4 and R6 are each independently selected from alkyl, alkoxy, halide, aminoalkyl, dialkylaminoalkyl, in which the alkyl and alkoxy groups contain from 1 to 10, preferably 1 to 6 carbons, and are straight or branched chains;
(v) any two of R3, R4, R5, R6, and R7, which are selected as in (i), form fused carbocyclic or heterocyclic rings.
In more preferred embodiments the above compounds in which, R2 is H, CH3, C2H5, or CF3; R1 is Cl, Br or CH3; n is 0 or 1; and R3, R4, R5, R6, R7, are selected from either (i), (ii), (iii) or (iv) as follows:
(i) R5 and R6 are H; R4 and R7 are each independently selected from H, halide, NH2, CF3, Ph, CH3; and R3 is selected from H, NHOH, NH2, EtNH, (CH3)2N, Phxe2x80x94CH2NH, NO2, F; Cl, Br, I, CN, CH3, (CH3)3C, C5H11, CH3O, n-C4H9O, CH2xe2x95x90CH, Phxe2x80x94CHxe2x95x90CH, CHxe2x89xa1C, Phxe2x80x94Cxe2x89xa1C, Ph, 3-(ethyoxycarbonylmethyl)ureido, and 3-cyclohexylureido; or
(ii) R4 and R7 together form 1,3-butadienyl, 4-chloro-1,3-butadienyl, 4-dimethylamino-1,3-butadienyl or 1-aza-1,3-butadienyl; and R3, R5 and R6 are defined as in (i) of this embodiment; or
(iii) R7 and R3 together form 1,3-butadienyl, 3-chloro-1,3-butadienyl, 4-dimethylamino-1,3-butadienyl or 1-aza-1,3-butadienyl; and R4, R5 and R6 are as defined in (i) of this embodiment; or
(iv) R3, R5, and R7 are H or as defined in (i); and R4 and R6 are each independently selected from alkyl, alkoxy, halide, amino alkyl, alkylaminoalkyl or dialkylaminoalkyl, in which the alkyl and alkoxy groups contain from 1 to 10, preferably 1 to 6 carbons, and are straight or branched chains are provided.
More preferred among the above compounds are those in which Ar2 is a substituted or unsubstituted phenyl, particularly biphenyl, or naphthyl; R1 is Br, Cl or I; R2 is H, CH3, C2H5, CF3, C2F5, n-C3H7, cycloC3H5, and C4H9; and R3, R4, R51, R6 and R7 are either (i), (ii), (iii), (iv) or (v):
(i) R5, R6 and R7 are H; n is 0 and R3 is H, NH2, CH3 CF3, halide, C2H5NH or Ph, R4 is H, CF3, NH2, R7 is H or CF3, and R5 and R6 are H; or
(ii) R3, R5 and R6 are H; n is 0 and R4 and R7 together form 1,3-butadienyl, 4-dimethylamino-1,3-butadienyl, 1-chloro-1,3-butadienyl, or 4-chloro-1,3-butadienyl; or
(iii) R4, R5 and R6 are H; n is 0; and R7 and R3 together form 1,3-butadienyl, 4-dimethylamino-1,3-butadienyl, 1-chloro-1,3-butadienyl, 1-aza-1,3-butadienyl; or
(iv) R4 is H or NH2, R5 and R6 are H; n is 1 and R3 is H, NH2 and halide; CH3, Br, Cl, F, CF3, NH2, R7 is H, CH3, Br, Cl, F, NH2 or CF3, and R5 and R6 are H; or
(v) R3, R5, and R7 are H are as defined in (i); and R4 and R6 are each independently selected from alkyl groups that contain from 1 to 6 carbons, and are straight or branched chains, lower alkoxy, and halide.
In more preferred embodiments, the benzenesulfonamides and naphthalenesulfonamdies are N-(4-halo)-substituted N-isoxazolylsulfonamides or are 4-higher alkyl-substituted N-isoxazolylsulfonamides, in which R2 is H, CH3, C2H5, C2F5 or CF3; and R3, R4, R5, R6 and R7 are either (i) or (ii) as follows:
(i) R4, R5, R6 and R7 are each independently selected from H, halide, NH2, CF3, Ph and CH3; R3 is selected from H, NHOH, NH2, C2H5NH, (CH3)2N, Phxe2x80x94CH2NH, NO2, F, Cl, Br, I, CN, CH3, (CH3)3C, C5H11, CH3O, n-C4H9O, CH2xe2x95x90CH, Phxe2x80x94CHxe2x95x90CH, CHxe2x89xa1C, Phxe2x80x94Cxe2x80x94C, Ph, 3-(ethyoxycarbonylmethyl)ureido, and 3-cyclohexylureido; or
(ii) R3, R5 and R7 are H; and R4 and R6 are each an alkyl group that contains from 1 to 3 carbons, which are straight or branched chains.
In yet more preferred embodiments, R1 is most preferably Br or Cl or higher alkyl (C9-C13); R2 is CH3, C2H5 or CF3; and R3, R4, R6 and R7 are (i) or (ii) as follows:
(i) R3 is H, NH2, CH3CF3, halide or C2H5NH; R4, R5 and R6 are independently selected from H, CF3, halide, particularly Br and Cl, NH2; and R7 is H, CH3, CH2CH3, (CH3)2CH, F or CF3; or
(ii) R3, R5 and R7 and R4 and R6 are each an methyl or ethyl.
In all embodiments, R1 is most preferably Br, except in instances in which enhanced ETB affinity, compared to the corresponding compound in which R1 is CH3, is desired, than R1 is most preferably a higher alkyl (8 to 15 carbons, preferably 9 to 13 carbons).
1. Compounds in Which Ar2 is Phenyl and Biphenyl and n is 0
Compounds in which Ar2 is phenyl or biphenyl have the following formulae (IV): 
in which R3, R4, R5, R6, and R7 are selected from (i) or (ii) with the proviso that, (a) when Ar2 is phenyl, R3 is not NO2, NH2, or lower alkyl, unless R1 is higher alkyl (C8-C15, preferably C9-C13), and (b) when Ar2 is naphthyl, 2-biphenyl, or phenyl, other than 3- or 4-biphenyl in which R4 and R5 is H, dibenzofuryl, dibenzothiophenyl or dibenzopyrrolyl, R1 is halide or higher alkyl:
(i) R3, R4, R5, R6, and R7 are each selected independently from among H, NHOH, NH2, NO2, N3, halide, pseudohalide, alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkoxy, alkylamino, alkylthio, alkoxyalkyl, alkylsulfinyl, alkylsulfonyl, aryloxy, arylamino, arylthio, arylsulfinyl, arylsulfonyl, haloalkyl, haloaryl, alkoxycarbonyl, alkylcarbonyl, arylcarbonyl, formyl, substituted or unsubstituted amido, substituted or unsubstituted ureido, where the alkyl, alkenyl, alkynyl portions are straight or branched chains of from about 1 up to about 10 carbons, preferably, 1 to about 5 or 6 carbons and the aryl portions contain from 3 up to about 10 carbons, preferably 6 carbons; are each independently selected as described above; or, alternatively,
(ii) R3, R5, and R7 are H; and R4 and R6 are each independently selected from alkyl, alkoxy, halide aminoalkyl, dialkylaminoalkyl, which are unsubstituted or substituted with alkyl groups, wherein the alkyl and alkoxy groups contain from 1 to 10, preferably 1 to 6 carbons, and are straight or branched chains.
Among the above phenyl and biphenyl compounds, are compounds with the following formulae (V): 
in which R3, R5 and R7 are each independently
(a) hydrogen, except that at least one of R3, R5 and R7 is other than hydrogen and any substituent at the 4-position on the phenyl ring is not NH2, NO2 or lower alkyl, when the other substituents are hydrogen;
(b) alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, aryl, aryloxy, aralkyl, or aralkoxy, any of which may be substituted with W1, W2 and W3, except that if one of R3, R5 are R7 is alkyl at the 4 position, at least one of the other two of R3, R5 are R7 is not hydrogen;
(c) halo;
(d) hydroxyl;
(e) cyano;
(f) nitro, except that if one of R3, R5 and R7 is 4-NO2, then at least one of the other two of R3, R5 and R7is not hydrogen;
(g) xe2x80x94C(O)H or xe2x80x94C(O)R27;
(h) xe2x80x94CO2H or xe2x80x94CO2R27;
(i) xe2x80x94SH, xe2x80x94S(O)nR27, xe2x80x94S(O)mxe2x80x94OH, xe2x80x94S(O)mOR27, xe2x80x94Oxe2x80x94S(O)mOH, or xe2x80x94Oxe2x80x94S(O)mOR27;
(j) xe2x80x94W4NR28R29, except that, if one of R3, R5 and R7 is 4-W4NR28R29 then at least one of the other two of R3, R5 and R 7is not hydrogen; or
(k) xe2x80x94W4N(R32)xe2x80x94W5NR30R31;
R1 is halide or is higher alkyl (about 8 carbons up to about 15 carbons in the chain, preferably C9-C13), except when the compounds are 3- or 4-biphenyls in which the 2-substituent is hydrogen, then R1 is selected, independently from R2 from the substituents set forth for R2;
R2 is selected from:
(a) hydrogen;
(b) alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, aryl, aryloxy, aralkyl, or aralkoxy, any of which may be substituted with W1, W2 and W3;
(c) hydroxyl;
(d) cyano;
(e) nitro;
(f) xe2x80x94C(O)H or xe2x80x94C(O)R27;
(g) xe2x80x94CO2H or xe2x80x94CO2R27;
(h) xe2x80x94SH, xe2x80x94S(O)nR27, xe2x80x94S(O)mxe2x80x94OH, xe2x80x94S(O)mOR27, xe2x80x94Oxe2x80x94S(O)mxe2x80x94R27, xe2x80x94Oxe2x80x94S(O)mOH, or xe2x80x94Oxe2x80x94S(O)mxe2x80x94OR27;
(i) xe2x80x94W4xe2x80x94NR28R29; or
(j) xe2x80x94W4N(R32)xe2x80x94W5xe2x80x94NR3R31;
R27 is alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, aryl, or aralkyl, any of which may be substituted with W1, w2 and W3;
R28 is
(a) hydrogen;
(b) alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, aryl, or aralkyl, any of which may be substituted with W1, w2 and W3;
(c) cyano;
(d) hydroxyl;
(e) xe2x80x94C(O)H or xe2x80x94C(O)R27;
(f) xe2x80x94CO2R27;
(g) xe2x80x94SH, xe2x80x94S(O)nR27, xe2x80x94S(O)mxe2x80x94OH, xe2x80x94S(O)mxe2x80x94OR27, xe2x80x94Oxe2x80x94S(O)mxe2x80x94R27, xe2x80x94Oxe2x80x94S(O)mOH, or xe2x80x94Oxe2x80x94S(O)mxe2x80x94OR27 except when W4 is xe2x80x94S(O)nxe2x80x94;
R29 is
(a) hydrogen;
(b) xe2x80x94C(O)H or xe2x80x94C(O)R27, except when W4 is xe2x80x94C(O)xe2x80x94 and R28 is xe2x80x94C(O)H, xe2x80x94C(O)R27, or xe2x80x94CO2R27;
(c) alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, aryl, or aralkyl, any of which may be substituted with W1, W2 and W3; or
R28 and R29 together are alkylene or alkenylene (either of which may be substituted with W1, W2 and W3), completing a 3- to 8-membered saturated, unsaturated or aromatic ring together with the nitrogen atom to which they are attached;
R30 is
(a) hydrogen;
(b) hydroxyl;
(c) xe2x80x94C(O)H or xe2x80x94C(O)R27;
(d) xe2x80x94CO2R27;
(e) xe2x80x94SH, xe2x80x94S(O)nR27, xe2x80x94S(O)mxe2x80x94OH, xe2x80x94S(O)mxe2x80x94OR27, xe2x80x94Oxe2x80x94S(O)mxe2x80x94R27, xe2x80x94Oxe2x80x94S(O)mOH, or xe2x80x94Oxe2x80x94S(O)mxe2x80x94OR27;
(f) alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, aryl, or aralkyl, any of which may be substituted with W1, W2 and W3;
R31 is
(a) hydrogen;
(b) xe2x80x94C(O)H or xe2x80x94C(O)R27, except when W5 is xe2x80x94C(O)xe2x80x94 and R30 is xe2x80x94C(O)H, xe2x80x94C(O)R27, or xe2x80x94CO2R27; or
(c) alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, aryl, or aralkyl, any of which may be substituted with W1, W2 and W3;
R32 is
(a) hydrogen;
(b) hydroxyl
(c) xe2x80x94C(O)H, xe2x80x94C(O)R27 or CO2R27; or
(d) alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, aryl, or aralkyl, any of which may be substituted with W1, W2 and W3;
or any two of R30, R31 and R32 together are alkylene or alkenylene (either of which may be substituted with W1, W2 and W3), completing a 3- to 8-membered saturated, unsaturated or aromatic ring together with the atoms to which they are attached; W1, W2 and W3 are each independently
(a) hydrogen;
(b) halo;
(c) hydroxy;
(d) alkyl;
(e) alkenyl;
(f) aralkyl;
(g) alkoxy;
(h) aryloxy;
(i) aralkoxy;
(j) xe2x80x94SH, xe2x80x94S(O)nW6, xe2x80x94S(O)mxe2x80x94OH, xe2x80x94S(O)mxe2x80x94OW6, xe2x80x94Oxe2x80x94S(O)mxe2x80x94W6, xe2x80x94Oxe2x80x94S(O)mOH, or xe2x80x94Oxe2x80x94S(O)mxe2x80x94OW6;
(k) oxo;
(l) nitro;
(m) cyano;
(n) xe2x80x94C(O)H or xe2x80x94C(O)W6;
(o) xe2x80x94CO2H or xe2x80x94CO2W6;
(p) xe2x80x94W4xe2x80x94NW7W8;
(q) W4xe2x80x94N(W11)xe2x80x94W5xe2x80x94W6; or
(r) xe2x80x94W4xe2x80x94N(W11)xe2x80x94W5xe2x80x94NW7W8;
W4 and W5 are each independently
(a) a single bond;
(b) xe2x80x94W9xe2x80x94S(O)nxe2x80x94W10xe2x80x94;
(c) xe2x80x94W9xe2x80x94C(O)xe2x80x94W10xe2x80x94;
(d) xe2x80x94W9xe2x80x94C(S)xe2x80x94W10xe2x80x94;
(e) xe2x80x94W9xe2x80x94Oxe2x80x94W10xe2x80x94;
(f) xe2x80x94W9xe2x80x94Sxe2x80x94W10xe2x80x94; or
(g) xe2x80x94W9xe2x80x94Oxe2x80x94C(O)xe2x80x94W10xe2x80x94;
W6, W7 and W8 are each independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, aryl, or aralkyl, or W7 and W8 together are alkylene or alkenylene, completing a 3- to 8-membered saturated, unsaturated or aromatic ring together with the nitrogen atom to which they are attached;
W9 and W10 are each independently a single bond, alkylene, alkenylene, or alkynylene;
W11 is
(a) hydrogen;
(b) hydroxyl;
(c) xe2x80x94C(O)H, xe2x80x94C(O)W6 or xe2x80x94CO2W6;
(d) alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, aryl, or aralkyl;
or any two of W7 and W8 and W11 together are alkylene or alkenylene, completing a 3- to 8-membered saturated, unsaturated, or aromatic ring together with the atoms to which they are attached;
m is 1 or 2; and
n is 0, 1, or 2.
Preferred compounds include those in which one of R3, R5 or R7 is phenyl or phenoxy or compounds in which one of R3, R5 or R7 is hydrogen, one of the other two of R3, R5 and R7 is at the 2 position and is not hydrogen, and the other of R3, R5 and R7 is at the 5 position. Thus, preferred compounds are 2-substituted benzenesulfonamides, and 2,5-substituted benzenesulfonamides. In addition, in preferred compounds R1 is preferably halide or higher alkyl. Preferred substituents are lower alkyl, particular methyl, ethyl, and propyl, halide, amino, dimethylamino, and methoxy.
(a) Benzenesulfonamides (Other than Biphenylsulfonamides)
Benzene sulfonamides are provided. In these compounds Ar2 has the formula 
in which:
R1 is halide or higher alkyl (greater than 8 carbons); R2 selected from H, NH2, NO2, halide, pseudohalide, alkyl, alkenyl, alkynyl, aryl, arylalkyl, heteroaryl, alkoxy, alkylamino, alkylthio, haloalkoxy, haloalkyl, alkylsulfinyl, alkylsulfonyl, aryloxy, arylamino, arylthio, arylsulfinyl, arylsulfonyl, aminocarbonyl, haloalkyl, haloaryl, alkoxycarbonyl, alkylcarbonyl, arylcarbonyl, formyl, substituted or unsubstituted amido, substituted or unsubstituted ureido, in which the alkyl, alkenyl and alkynyl portions are either straight or branched chains that contain from 1 up to about 10 carbon atoms, and the aryl portions contain from about 4 to about 14 carbons; and
R3, R4, R5, R6, and R7 are either (i) or (ii) as follows:
(i) R3, R4, R5, R6, and R7 are selected independently from among H, NHOH, NH2, NO2, pseudohalide, including N3, halide, alkenyl, alkynyl, aryl, heteroaryl, alkoxy, alkylamino, alkylthio, alkoxyalkyl, alkylsulfinyl, alkylsulfonyl, aryloxy, arylamino, arylthio, arylsulfinyl, arylsulfonyl, haloalkyl, haloaryl, alkoxycarbonyl, alkylcarbonyl, arylcarbonyl, formyl, substituted or unsubstituted amido, substituted or unsubstituted ureido, where the alkyl, alkenyl, alkynyl portions are straight or branched chains of from about 1 up to about 10 carbons, preferably, 1 to about 5 or 6 carbons, are unsubstituted or substituted with groups set forth for Z, above, and the aryl portions contain from 3 up to about 10 carbons, preferably 3 to 6 carbons, and, also are unsubstituted or substituted with groups, independently selected from Z, with the proviso that, if R4, R5, R6, and R7 are all hydrogen, then R3is not NO2, NH2 or lower alkyl; or
(ii) R3, R5, and R7 are H; and R4 and R6 are each independently selected from alkyl, alkoxy, halide, aminoalkyl, dialkylamino, dialkylaminoalkyl, which are unsubstituted or substituted with alkyl groups, and in which the alkyl and alkoxy groups contain from 1 to 10, preferably 1 to 6 carbons, and are straight or branched chains.
Compounds in which at least one of R3-R7 is phenyl are discussed below with the biphenyl compounds.
In certain preferred embodiments: R1 is halide or a higher alkyl selected from C9H19 to C13H27; R2 is selected independently from alkyl, lower alkenyl, lower alkynyl, lower haloalkyl and H; and R3, R4, R5, R6, and R7 are either (i) or (ii) as follows:
(i) R4, R5, R6 and R7 are each independently selected from H, lower alkyl, NH2, NO2, halide, pseudohalide; R3 is selected from H, NHOH, halide, pseudohalide, alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkoxy, alkylamino, alkylthio, alkoxyalkyl, alkylsulfinyl, alkylsulfonyl, aryloxy, arylamino, arylthio, arylsulfinyl, arylsulfonyl, haloalkyl, haloaryl, alkoxycarbonyl, alkylcarbonyl, arylcarbonyl, formyl, substituted or unsubstituted amido, substituted or unsubstituted ureido, where the alkyl, alkenyl, alkynyl portions are straight or branched chains of from 1 up to 5 or 6 carbons and the aryl portions contain from 4 to 14 carbons; or
(ii) R3, R5, and R7 are H; and R4 and R6 are each independently selected from alkyl, alkoxy, halide, aminoalkyl, and dialkylaminoalkyl, which are unsubstituted or substituted with alkyl groups, and in which the alkyl and alkoxy groups contain 1 to 6 carbons, and are straight or branched chains.
In more preferred embodiments, R1 is Cl or Br, or if greater ETB activity is preferred a higher alkyl (C9H19 to C13H27; R2 is selected from H, CH3, C2H5, CF3, C2F5, n-C3H7, cyclo-C3H7, nC13H27 and nC9H19; and R3, R4, R5, R6, and R7 are either (i) or (ii) as follows:
(i) R4, R5, R6 and R7 are each independently selected from H, halide, NH2, CF3, Ph and CH3; R3 is selected from H, NHOH, NH2, C2H5NH, (CH3)2N, Phxe2x80x94CH2NH, NO2, F, Cl, Br, I, CN, CH3, (CH3)3C, C5H11, CH3O, n-C4H9O, CH2xe2x95x90CH, Phxe2x80x94CHxe2x95x90CH, CHxe2x89xa1C, Phxe2x80x94Cxe2x89xa1C, Ph, 3-(ethyoxycarbonylmethyl)ureido, and 3-cyclohexylureido; or
(ii) R3, R5, and R7 are H; and R4 and R6 are each independently selected from alkyl and aminoalkyl in which the alkyl groups have from 1 to 6 carbons that may from straight or branched chains.
In yet more preferred embodiments, R1 is Br, Cl or C9H19 to C13H27; R2 is H, CH3, C2H5, or CF3; and R3, R4, R5, R6, and R7 are either (i) or (ii) as follows:
(i) R3 is H, NH2, CH3 CF3, halide or C2H5NH; R4, R5 and R6 are independently selected from H, CH3, C2H5, (CH3)2CH, CF3, halide, particularly Br and Cl, NH2; and R7 is H, CH3, CH2CH3, (CH3)2CH, F or CF3; or
(ii) R3, R5, and R7 are H; and R4 and R6 are each independently selected from alkyl groups in which the alkyl groups have from 1 to 3 carbons and may form straight or branched chains.
Of the above compounds those with ortho and/or meta substituents or those that are substituted at positions 2 and 5 on the benzene ring are generally more preferred, except when the resulting compound is a biphenyl and ETB affinity is desired, then the corresponding para-substituted compounds are preferred. Compounds with ortho substituents are more generally more preferred than the corresponding meta-substituted compounds. This observation is particularly important when activity with respect to ETA receptors is considered. In addition, in preferred compounds R1 is preferably halide. Preferred substituents are lower alkyl, particular methyl, ethyl, and propyl, halide, amino, dimethylamino, and methoxy. Other preferred substituents may be deduced from the following Table.
Benzenesulfonamides were synthesized and tested using the exemplified assays (see, EXAMPLES) and selected results are set forth in Table 1 (the N-(3,4-dimethyl-5-isoxazolyl)benzenesulfonamedes, which are not included in the above formulae, are included for comparison with the corresponding N-94-halo-3-methyl-5-isoxazolyl)benzenesulfonamide.
(b) Ar2 is biphenyl
Compounds of formulae I and II in which Ar1 is N-(5-isoxazolyl) or N-(3-isoxazolyl) in which Ar2 is selected from biphenyl or styryl derivatives. These compounds can be represented by the following formulae (III): 
in which n is 0 to 10, preferably 0 to 6, more preferably 0 to 3;
R1 and R2 are either (i), (ii) or (iii) as follows:
(i) R1 and R2 are each independently selected from H, NH2, NO2, halide, pseudohalide, alkyl, alkenyl, alkynyl, aryl, arylalkyl, heteroaryl, alkoxy, alkylamino, alkylthio, alkyloxy, haloalkyl, alkylsulfinyl, alkylsulfonyl, aryloxy, arylamino, arylthio, arylsulfinyl, arylsulfonyl, haloalkyl, haloaryl, alkoxycarbonyl, alkylcarbonyl, aminocarbonyl, arylcarbonyl, formyl, substituted or unsubstituted amido, substituted or unsubstituted ureido, in which the alkyl, alkenyl and alkynyl portions contain from 1 up to about 14 carbon atoms and are either straight or branched chains or cyclic, and the aryl portions contain from about 4 to about 16 carbons, except that R2 is not halide or pseudohalide; or,
(ii) R1 and R2 together form xe2x80x94(CH2)n, where n is 3 to 6; or,
(iii) R1 and R2 together form 1,3-butadienyl, and with the proviso that Ar2 is not 2-biphenyl or biphenyl in which R4 and/or R5 is other than hydrogen unless R1 is a halide or higher (C8-C15, preferably C9-C13) alkyl;
at least one of R3, R4, R5, R6, and R7 is phenyl, and the remaining of R3, R4, R5, R6 and R7 are selected with the proviso that: (a) when one of R3, R6, and R7 is phenyl, then R4 and R5 are hydrogen, unless R1 is halide or higher alkyl (C8-C15, preferably C9-C13), and (b) when one of R4 or R5 is phenyl, then the other is hydrogen, unless R1 is halide or higher alkyl (C8-C15, preferably C9-C13):
the others of R3, R4, R5, R6, and R7 are selected independently from among H, NHOH, NH2, NO2, pseudohalide, including N3, halide, alkenyl, alkynyl, aryl, heteroaryl, alkoxy, alkylamino, alkylthio, alkoxyalkyl, alkylsulfinyl, alkylsulfonyl, aryloxy, arylamino, arylthio, arylsulfinyl, arylsulfonyl, haloalkyl, haloaryl, alkoxycarbonyl, alkylcarbonyl, arylcarbonyl, formyl, substituted or unsubstituted amido, substituted or unsubstituted ureido, where the alkyl, alkenyl, alkynyl portions are straight or branched chains of from about 1 up to about 10 carbons, preferably, 1 to about 5 or 6 carbons, are unsubstituted or substituted with Z, above, and the aryl portions contain from 3 up to about 10 carbons, preferably 3 to 6 carbons, and, also are unsubstituted or substituted with groups
In more preferred embodiments, R1 is Cl or Br, or if greater ETB activity is preferred a higher alkyl (C8H19 to C13H27; R2 is selected from H, CH3, C2H5, CF3, C2F5, n-C3H7, cyclo-C3H7, nC13H27 and nC9H19; and R3, R4, R5, R6, and R7 are either (i) or (ii) as follows:
(i) R4, R5, R6 and R7 are each independently selected from H, halide, NH2, CF3, Ph and CH3; R3 is selected from H, NHOH, NH2, C2H5NH, (CH3)2N, Phxe2x80x94CH2NH, N2, F, Cl, Br, I, CN, CH3, (CH3)3C, C5H11, CH3O, n-C4H9O, CH2xe2x95x90CH, Phxe2x80x94CHxe2x95x90CH, CHxe2x89xa1xe2x80x94C, Phxe2x80x94Cxe2x89xa1C, Ph, 3-(ethyoxycarbonylmethyl)ureido, and 3-cyclohexylureido; or
(ii) R3, R5, and R7 are H; and one of R4 and R6 is phenyl and the other is selected from alkyl and aminoalkyl in which the alkyl groups have from 1 to 6 carbons that may form straight or branched chains.
In yet more preferred embodiments, R1 is Br, Cl or C9H19 to C13H27; R2 is H, CH3, C2H5, or CF3; and R3, R4, R5, R6, and R7 are either (i) or (ii), with the proviso that at least one of them is substituted or unsubstituted phenyl, as follows:
(i) R3 is H, NH2, CH3 CF3, Ph, halide or C2H5NH; R4, R1 and R6 are independently selected from H, CH3, C2H5, (CH3)2CH, CF3, halide, particularly Br and Cl, Ph and NH2; and R7 is H, CH3, CH2CH3, (CH3)2CH, F or CF3; or
(ii) R3, R5, and R7 are H or phenyl; and R4 and R6 are each independently selected from alkyl groups in which the alkyl groups have from 1 to 3 carbons and may form straight or branched chains.
In yet more preferred embodiments, R1 is Br, Cl or C9H19 to C13H27; R2 is H, CH3, C2H5, or CF3; and R3, R4, R5, R6, and R7 are, with the proviso that at one of R3, R4, R5, R6, and R7 is unsubstituted or substituted phenyl, selected as follows:
R3 is Ph, H, NH2, CH3 CF3, halide or C2H5NH; R4, R5 and R6 are independently selected from Ph, H, CH3, C2H5, (CH3)2CH, CF3, halide, particularly Br and Cl, NH2; and R7 is Ph, H, CH3, CH2CH3, (CH3)2CH, F or CF3.
For the 3- or 4-biphenyl compounds, in which R4 and/or R5 is other than hydrogen, and for the 2-biphenyl compounds, R1 is preferably bromide or chloride, methyl or ethyl or (C9-C13)alkyl, and R2 is preferably alkyl. In the most active compounds provided herein, as evidenced by in vitro binding assays, R1 is bromide or chloride or, in instances in which ETB selectivity is desired, C9-C13 alkyl. R1 is preferably halide, lower alkyl, particularly CH3, or C9H19-C13H27.
In preferred embodiments, in which the sulfonamides are biphenylsulfonamides in which R1 is halide; R2 is selected from alkyl, lower alkenyl, lower alkynyl, lower haloalkyl and H; and the others of R3-R7 that are not phenyl are selected from H, lower alkyl, haloalkyl and halide. In preferred of these embodiments, R1 is Cl or Br, and for the 3-biphenylsulfonamides and, particularly, the 4-biphenylsulfonamides, R1 is also CH3; R2 is selected from H, CH3, C2H5, CF3, C2F5, n-C3H7 and cyclo-C3H7; and R26 and R13 are each independently selected from H, halide, NH2, CF3 CH3, CN, CH3, (CH3)3C, C5H11, CH3O, n-C4H9O and CH2xe2x95x90CH.
In yet more preferred embodiments, R2 is H, CH3, C2H5, or CF3; R3-R7 are independently selected from H, CH3, C2H5, CF3, and halide; and X is O.
In another preferred embodiment, the biphenylsulfonamides are 3- or 4-biphenylsulfonamides, in which R4 and R1 are other than hydrogen.
In such instances R1 is preferably, halide or methyl or higher (C9-C13) alkyl. Such compounds have a higher ETB affinity than the 2-biphenylsulfonamides. It is also preferred that the substituent at the 2-position is hydrogen. R1 is selected from halide, CH3, C2H5, CF3, C2F5, n-C3H7 and cyclo-C3H7, preferably halide or CH3, and R2 is selected from H, CH3, C2H5, CF3, C2F5, n-C3H7 and cyclo-C3H7; and R3-R1 are each independently selected from H, halide, NH2, CF3 CH3, CN, CH3, (CH3)3C, C5H11, CH3O, n-C4H9O and CH2xe2x95x90CH. In more preferred of these embodiments, R1 is halide or CH3, and R2 are selected from H, CH3, C2H5, or CF3; R3-R7 are independently selected from H, CH3, C2H5, CF3, and halide.
When Ar2 is a biphenyl (n=0) in which R4 and R5 are other than hydrogen, the compounds are (4-halo-isoxazoly)sulfonamides or are (4-higher alkyl-isoxazoly)sulfonamides, in which the alkyl group contains more than about 8, preferably 9 to 15, more preferably 9 to 13, carbons atoms.
In embodiments described in detail herein, preferred compounds include compounds that are ETB receptor selective or that bind to ETB receptors with an IC50 of less than about 1 xcexcM (when measured in the assays herein at 4xc2x0 C.). In these compounds, Ar2 is 3-biphenyl, 4-biphenyl and R1 is halide or preferably higher alkyl (C9H19 to C13H27). R2 is selected from among alkyl, lower haloalkyl, H; and R1 is halide, lower alkyl or lower haloalkyl, or, when Ar2 is phenyl or naphthyl, R1 is higher alkyl (nine or more carbon atoms, preferably 9 to 13 carbon atoms). When R4 or R5 are other than hydrogen, then Ar1 is 4-haloisoxazolyl or 4-higher (C8 to C15, preferably C9H19 to C13H27) alkylisoxzaolyl are preferred.
Among the above phenyl and biphenyl compounds, are compounds with the following formulae (V): 
in which R3, R5 and R7 are each independently selected as follows, with the proviso that at least one of R3, R5 and R7 is phenyl or phenoxy:
(a) hydrogen;
(b) alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, aryl, aryloxy, aralkyl, or aralkoxy, any of which may be substituted with W1, W2 and W3;
(c) halo;
(d) hydroxyl;
(e) cyano;
(f) nitro;
(g) xe2x80x94C(O)H or xe2x80x94C(O)R27;
(h) xe2x80x94CQ2H or xe2x80x94CO2R27;
(i) xe2x80x94SH, xe2x80x94S(O)nR27, xe2x80x94S(O)mxe2x80x94OH, xe2x80x94S(O)mxe2x80x94OR27, Oxe2x80x94S(O)mOH, or xe2x80x94Oxe2x80x94S(O)mOR27;
(j) xe2x80x94W4NR28R29, except that, if one of R3, R5 and R7 is 4-W4NR28R29 then at least one of the other two of R3, R5 and R7is not hydrogen; or
(k) xe2x80x94W4N(R32)xe2x80x94W5NR30R31;
R1 and R2 are each independently selected from:
(a) hydrogen;
(b) alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, aryl, aryloxy, aralkyl, or aralkoxy, any of which may be substituted with W1, W2 and W3;
(c) hydroxyl;
(d) cyano;
(e) nitro;
(f) xe2x80x94C(O)H or xe2x80x94C(O)R27;
(g) xe2x80x94CO2H or xe2x80x94CO2R27;
(h) xe2x80x94SH, xe2x80x94S(O)nR27, xe2x80x94S(O)mxe2x80x94OH, xe2x80x94S(O)mOR27, xe2x80x94Oxe2x80x94S(O)mxe2x80x94R27, xe2x80x94O""S(O)mOH, or xe2x80x94Oxe2x80x94S(O)mxe2x80x94OR27;
(i) xe2x80x94W4xe2x80x94NR28R29; or
(j) xe2x80x94W4N(R32)xe2x80x94W5xe2x80x94NR30R31;
R1 is selected with the proviso that when Ar2 is a 3- or 4-biphenyl in which the substituent at the 2-position is other than hydrogen or when the compounds are 2-biphenyls, then R1 is halide or higher allyl (C8H17 to C15H31, preferably C9H19 to C13H27);
R27 is alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, aryl, or aralkyl, any of which may be substituted with W1, W2 and W3;
R28 is
(a) hydrogen;
(b) alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, aryl, or aralkyl, any of which may be substituted with W1, W2 and W3;
(c) cyano;
(d) hydroxyl;
(e) xe2x80x94C(O)H or xe2x80x94C(O)R27;
(f) xe2x80x94CO2R27;
(g) xe2x80x94SH, xe2x80x94S(O)R27, xe2x80x94S(O)mxe2x80x94OH, xe2x80x94S(O)mxe2x80x94OR27, xe2x80x94S(O)mxe2x80x94R27, xe2x80x94S(O)mOH, or xe2x80x94Oxe2x80x94S(O)mxe2x80x94OR27, except when W4 is xe2x80x94S(O)nxe2x88x92;
R29 is
(a) hydrogen;
(b) xe2x80x94C(O)H or xe2x80x94C(O)R27, except when W4 is xe2x80x94C(O)xe2x80x94 and R28 is xe2x80x94C(O)H, xe2x80x94C(O)R27, or xe2x80x94CO2R27;
(c) alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, aryl, or aralkyl, any of which may be substituted with W1, W2 and W3; or
R28 and R29 together are alkylene or alkenylene (either of which may be substituted with W1, W2 and W3), completing a 3- to 8-membered saturated, unsaturated or aromatic ring together with the nitrogen atom to which they are attached;
R30 is
(a) hydrogen;
(b) hydroxyl;
(c) xe2x80x94C(O)H or xe2x80x94C(O)R27;
(d) xe2x80x94CO2R27;
(e) xe2x80x94SH, xe2x80x94S(O)nR27, xe2x80x94S(O)mxe2x80x94OH, xe2x80x94S(O)mxe2x80x94OR27, xe2x80x94Oxe2x80x94S(O)mR27, xe2x80x94Oxe2x80x94S(O)mOH, or xe2x80x94Oxe2x80x94S(O)mxe2x80x94OR27;
(f) alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, aryl, or aralkyl, any of which may be substituted with W1, W2 and W3,
R31 is
(a) hydrogen;
(b) xe2x80x94C(O)H or xe2x80x94C(O)R27, except when W5 is xe2x80x94C(O)xe2x80x94 and R30 is xe2x80x94C(O)H, xe2x80x94C(O)R27, or xe2x80x94CO2R27; or
(c) alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, aryl, or aralkyl, any of which may be substituted with W1, W2 and W3;
R32 is
(a) hydrogen;
(b) hydroxyl
(c) xe2x80x94C(O)H, xe2x80x94C(O)R27 or CO2R27; or
(d) alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, aryl, or aralkyl, any of which may be substituted with W1, W2 and W3;
or any two of R30, R31 and R32 together are alkylene or alkenylene (either of which may be substituted with W1, W2 and W3), completing a 3- to 8-membered saturated, unsaturated or aromatic ring together with the atoms to which they are attached; W1, W2 and W3 are each independently
(a) hydrogen;
(b) halo;
(c) hydroxy;
(d) alkyl;
(e) alkenyl;
(f) aralkyl;
(g) alkoxy;
(h) aryloxy;
(i) aralkoxy;
(j) xe2x80x94SH, xe2x80x94S(O)nW6, xe2x80x94S(O)mxe2x80x94OH, xe2x80x94S(O)mxe2x80x94OW6, xe2x80x94Oxe2x80x94S(O)mxe2x80x94W6, xe2x80x94Oxe2x80x94S(O)mOH, or xe2x80x94Oxe2x80x94S(O)mxe2x80x94OW6;
(k) oxo;
(l) nitro;
(m) cyano;
(n) xe2x80x94C(O)H or xe2x80x94C(O)W6;
(o) xe2x80x94CO2H or xe2x80x94CO2W6;
(p) xe2x80x94W4xe2x80x94NW7W8;
(q) W4xe2x80x94N(W11)xe2x80x94W5xe2x80x94W6; or
(r) xe2x80x94W4xe2x80x94N(W11)xe2x80x94W5xe2x80x94NW7W8;
W4 and W5 are each independently
(a) a single bond;
(b) xe2x80x94W9xe2x80x94S(O)nxe2x80x94W10xe2x80x94;
(c) xe2x80x94W9xe2x80x94C(O)xe2x80x94W10xe2x80x94;
(d) xe2x80x94W9xe2x80x94C(S)xe2x80x94W10xe2x80x94;
(e) xe2x80x94W9xe2x80x94Oxe2x80x94W10xe2x80x94;
(f) xe2x80x94W9xe2x80x94Sxe2x80x94W10xe2x80x94; or
(g) xe2x80x94W9xe2x80x94Oxe2x80x94C(O)xe2x80x94W10xe2x80x94;
W6, W7 and W8 are each independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, aryl, or aralkyl, or W7 and W8 together are alkylene or alkenylene, completing a 3- to 8-membered saturated, unsaturated or aromatic ring together with the nitrogen atom to which they are attached;
W9 and W10 are each independently a single bond, alkylene, alkenylene, or alkynylene;
W11 is
(a) hydrogen;
(b) hydroxyl;
(c) xe2x80x94C(O)H, xe2x80x94C(O)W6 or xe2x80x94CO2W6;
(d) alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, aryl, or aralkyl;
or any two of W7 and W8 and W11 together are alkylene or alkenylene, completing a 3- to 8-membered saturated, unsaturated, or aromatic ring together with the atoms to which they are attached;
m is 1 or 2; and
n is 0, 1, or 2,
with the proviso that (i) R2 is not halide; and (ii) unless R5 is hydrogen, R1 is halide or is higher alkyl (greater than about 8 carbons up to 15 carbons in the chain, preferably nine to thirteen carbons).
If R5 is hydrogen, then R1 is also preferably selected from lower alkyl, preferably methyl or ethyl, halide or higher alkyl.
Preferred compounds among these include those in which one of R3, R5 or R1 is phenyl. Preferred substituents on the rings are lower alkyl, particular methyl, ethyl, and propyl, halide, amino, dimethylamino, and methoxy. In addition, in preferred compounds R1 is preferably halide or higher alkyl.
In more preferred compounds, R1 is Cl or Br, or if greater ETB activity is preferred a higher alkyl (C9H19 to C13H27; R2 is selected from H, CH3, C2H5, CF3, C2F5, n-C3H7, cyclo-C3H7, nC13H27 and nC9H19; and R3, R3, R5, and R7 are each independently selected from H, halide, NH2, CF3, Ph and CH3; R3 is selected from H, NHOH, NH2, C2H5NH, (CH3)2N, Phxe2x80x94CH2NH, NO2, F, Cl, Br, I, CN, CH3, (CH3)3C, C5H11, CH3O, n-C4H9O, CH2xe2x95x90CH, Phxe2x80x94CHxe2x95x90CH, CHxe2x89xa1C, Phxe2x80x94Cxe2x89xa1C, Ph, 3-(ethyoxycarbonylmethyl)ureido, and 3-cyclohexylureido, with the proviso that at least one of R3, R5, and R7 is a substituted or unsubstituted phenyl. Preferred substituents are lower alkyl, alkoxy(lower)alkyl, halide, and the like. Preferred positions for the substituents are ortho and/or para.
In yet more preferred embodiments, R1 is Br, Cl or C9H19 to C13H27; R2 is H, CH3, C2H5, or CF3; and R3, R5 and R7, with the proviso that at least one of R3, R5 and R7 is phenyl, are selected as follows:
R3 is H, Ph, NH2, CH3 CF3, halide or C2H5NH; R5 is selected from H, CH3, C2H5, (CH3)2CH, CF3, halide, particularly Br and Cl, NH2 or Ph; and R7 is H, Ph, CH3, CH2CH3, (CH3)2CH, F or CF3.
Of any of these compounds described above, those with formulae VI: 
in which R5 is hydrogen are more preferred. In these compounds R3 and R7 are each independently selected as follows, with the proviso that one of R3 and R7 is phenyl or phenoxy, preferably phenyl and the other is:
(a) hydrogen;
(b) alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, aryl, aryloxy, aralkyl, or aralkoxy, any of which may be substituted with W1, W2 and W3;
(c) halo;
(d) hydroxyl;
(e) cyano;
(f) nitro;
(g) xe2x80x94C(O)H or xe2x80x94C(O)R27;
(h) xe2x80x94CO2H or xe2x80x94CO2R27;
(i) xe2x80x94SH, xe2x80x94S(O)nR27, xe2x80x94S(O)mxe2x80x94OH, xe2x80x94S(O)mOR27, xe2x80x94S(O)mOH, or xe2x80x94Oxe2x80x94S(O)mOR27;
(j) xe2x80x94W4NR28R29, except that, if one of R3, R5 and R7 is 4xe2x80x94W4NR28R29 then at least one of the other two of R3, R5 and R7 is not hydrogen; or
(k) xe2x80x94W4N(R32)_W5NR30R31;
R1 and R2 are each independently selected from:
(a) hydrogen;
(b) alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, aryl, aryloxy, aralkyl, or aralkoxy, any of which may be substituted with W1, W2 and W3;
(c) hydroxyl;
(d) cyano;
(e) nitro;
(f) xe2x80x94C(O)H or xe2x80x94C(O)R27;
(g) xe2x80x94CO2H or xe2x80x94CO2R27;
(h) xe2x80x94SH, xe2x80x94S(O)nR27, xe2x80x94S(O)mxe2x80x94OH, xe2x80x94S(O)mOR27, xe2x80x94Oxe2x80x94S(O)mR27, xe2x80x94Oxe2x80x94S(O)mOH, or xe2x80x94Oxe2x80x94S(O)mxe2x80x94OR27;
(i) xe2x80x94W4xe2x80x94NR28R29; or
(j) xe2x80x94W4N(R32)xe2x80x94W5xe2x80x94NR30R31;
R27 is alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, aryl, or aralkyl, any of which may be substituted with W1, W2 and W3;
R28 is
(a) hydrogen;
(b) alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, aryl, or aralkyl, any of which may be substituted with W1, W2 and W3;
(c) cyano;
(d) hydroxyl;
(e) xe2x80x94C(O)H or xe2x80x94C(O)R27;
(f) xe2x80x94CO2R27;
(g) xe2x80x94SH, xe2x80x94S(O)nR27, xe2x80x94S(O)mxe2x80x94OH, xe2x80x94S(O)mxe2x80x94OR27, xe2x80x94Oxe2x80x94S(O)mR27, xe2x80x94Oxe2x80x94S(O)mOH, or xe2x80x94Oxe2x80x94S(O)mxe2x80x94OR27 except when W4 is xe2x80x94S(O)nxe2x80x94;
R29 is
(a) hydrogen;
(b) xe2x80x94C(O)H or xe2x80x94C(O)R27, except when W4 is xe2x80x94C(O)xe2x80x94 and R28 is xe2x80x94C(O)H, xe2x80x94C(O)R27, or xe2x80x94C2R27;
(c) alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, aryl, or aralkyl, any of which may be substituted with W1, W2 and W3; or
R28 and R29 together are alkylene or alkenylene (either of which may be substituted with W1, W2 and W3), completing a 3- to 8-membered saturated, unsaturated or aromatic ring together with the nitrogen atom to which they are attached;
R30 is
(a) hydrogen;
(b) hydroxyl;
(c) xe2x80x94C(O)H or xe2x80x94C(O)R27;
(d) xe2x80x94CO2R27;
(e) xe2x80x94SH, xe2x80x94S(O)nR27, xe2x80x94S(O)mxe2x80x94OH, xe2x80x94S(O)mxe2x80x94OR27 xe2x80x94Oxe2x80x94S(O)mxe2x80x94R27, xe2x80x94Oxe2x80x94S(O)mOH, or xe2x80x94Oxe2x80x94S(O)mxe2x80x94OR27;
(f) alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, aryl, or aralkyl, any of which may be substituted with W1, W2 and W3;
R31 is
(a) hydrogen;
(b) xe2x80x94C(O)H or xe2x80x94C(O)R27, except when W5 is xe2x80x94C(O)xe2x80x94 and R30 is xe2x80x94C(O)H, xe2x80x94C(O)R27, or xe2x80x94CO2R27; or
(c) alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, aryl, or aralkyl, any of which may be substituted with W1, W2 and W3;
R32 is
(a) hydrogen;
(b) hydroxyl
(c) xe2x80x94C(O)H, xe2x80x94C(O)R27 or CO2R27; or
(d) alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, aryl, or aralkyl, any of which may be substituted with W1, W2 and W3;
or any two of R30, R31 and R32 together are alkylene or alkenylene (either of which may be substituted with W1, W2 and W3), completing a 3- to 8-membered saturated, unsaturated or aromatic ring together with the atoms to which they are attached;
W1, W2 and W3 are each independently
(a) hydrogen;
(b) halo;
(c) hydroxy;
(d) alkyl;
(e) alkenyl;
(f) aralkyl;
(g) alkoxy;
(h) aryloxy;
(i) aralkoxy;
(j) xe2x80x94SH, xe2x80x94S(O)nW6, xe2x80x94S(O)mxe2x80x94OH, xe2x80x94S(O)mxe2x80x94OW6, xe2x80x94Oxe2x80x94S(O)mxe2x80x94W6, xe2x80x94Oxe2x80x94S(O)mOH, or xe2x80x94Oxe2x80x94S(O)mxe2x80x94OW6;
(k) oxo;
(l) nitro;
(m) cyano;
(n) xe2x80x94C(O)H or xe2x80x94C(O)W6;
(o) xe2x80x94CO2H or xe2x80x94CO2W6;
(p) xe2x80x94W4xe2x80x94NW7W8;
(q) W4xe2x80x94N(W11)xe2x80x94W5xe2x80x94W6; or
(r) xe2x80x94W4xe2x80x94N(W11)xe2x80x94W5xe2x80x94NW7W8;
W4 and W5 are each independently
(a) a single bond;
(b) xe2x80x94W9xe2x80x94S(O)nxe2x80x94W10xe2x80x94;
(c) xe2x80x94W9xe2x80x94C(O)xe2x80x94W10xe2x80x94;
(d) xe2x80x94W9xe2x80x94C(S)xe2x80x94W10xe2x80x94;
(e) xe2x80x94W9xe2x80x94Oxe2x80x94W10xe2x80x94;
(f) xe2x80x94W9xe2x80x94Sxe2x80x94W10xe2x80x94; or
(g) xe2x80x94W9xe2x80x94Oxe2x80x94C(O)xe2x80x94W10xe2x80x94;
W6, W7 and W8 are each independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, aryl, or aralkyl, or W7 and W8 together are alkylene or alkenylene, completing a 3- to 8-membered saturated, unsaturated or aromatic ring together with the nitrogen atom to which they are attached;
W9 and W10 are each independently a single bond, alkylene, alkenylene, or alkynylene;
W11 is
(a) hydrogen;
(b) hydroxyl;
(c) xe2x80x94C(O)H, xe2x80x94C(O)W6 or xe2x80x94CO2W6;
(d) alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, aryl, or aralkyl;
or any two of W7 and W8 and W11 together are alkylene or alkenylene, completing a 3- to 8-membered saturated, unsaturated, or aromatic ring together with the atoms to which they are attached;
m is 1 or 2; and
n is 0, 1, or 2.
In the more preferred of these compounds, R1, R2, R3 and R7 are selected as in the more preferred compounds of formulae (V). Most preferred are compounds in which Ar2 is 4-biphenyl.
The biphenyl compounds provided herein are generally ETB active or ETB selective (see, e,g., Table 2); i.e. the compounds provided herein inhibit binding of endothelin to ETB receptors at concentrations about 10- to about 30-fold less than they inhibit binding of endothelin to ETA receptors. In particular the 4-biphenylsulfonamides are ETB selective.
In general in all embodiments herein, 4-haloisoxazolyl sulfonamides exhibit substantially enhanced activity with respect to at least one of the ET receptors (about two-fold to twenty-fold greater activity), as assessed by assays, such as those provided herein, that measure binding to ETA and/or ETB receptors, compared to corresponding sulfonamides in which the substituent at the 4 position in the isoxazolyl is other than halo, such as alkyl. For example: the IC50 of N-(3,4-dimethyl-5-isoxazolyl)-2-biphenylsulfonamide for ETA receptors is about 0.008 xcexcM, whereas, the IC50 of N-(4-bromo-3-methyl-5-isoxazolyl)-2-biphenylsulfonamide is about 0.001 6 xcexcM (see, Table below); and (3) the IC50 of N-(3,4-dimethyl-5-isoxazolyl)-3-biphenylsulfonamide for ETB receptors is about 3.48 xcexcM; whereas, the IC50 of N-(4-bromo-3-methyl-5-isoxazolyl)-3-biphenylsulfonamide for ETB receptors is about 0.76 xcexcM and the IC50 of N-(4-chloro-3-methyl-5-isoxazolyl)-3-biphenylsulfonamide for ETB receptors is about 0.793 xcexcM (see, Table below).
Exemplary biphenyl sulfonamides are the following and those set forth in Table 2, and include, but are not limited to: N-(4-bromo-3-methyl-5-isoxazolyl)-4-biphenylsulfonamide, N-(4-chloro-3-methyl-5-isoxazolyl)-4-biphenylsulfonamide, N-(4-bromo-5-methyl-3-isoxazolyl)-4-biphenylsulfonamide, N-(4-chloro-5-methyl-3-isoxazolyl)-4-biphenylsulfonamide, N-(3-methyl-5-isoxazolyl)-4xe2x80x2-methylphenyl-4-biphenylsulfonamide, N-(4-bromo-3-methyl-5-isoxazolyl)-4xe2x80x2-methylphenyl-4-biphenylsulfonamide, N-(4-chloro-3-methyl-5-isoxazolyl)-4xe2x80x2-methylphenyl-4-biphenylsulfonamide, (3-methyl-5-isoxazolyl)-4xe2x80x2-trifluorophenyl-4-biphenylsulfonamide, (4-bromo-3-methyl-5-isoxazolyl)-4xe2x80x2-trifluorophenyl-4-biphenylsulfonamide, (3-methyl-5-isoxazolyl)-4xe2x80x2-methyoxyphenyl-4-biphenylsulfonamide, (4-bromo-3-methyl-5-isoxazolyl)-4xe2x80x2-methoxyphenyl-4-biphenylsulfonamide, (4-bromo-3-methyl-5-isoxazoly)-3xe2x80x2-methoxyphenyl-4-biphenylsulfonamide, (4-bromo-3-methyl-5-isoxazolyl)-2xe2x80x2-methoxyphenyl-4-biphenylsulfonamide, N-(4-bromo-3-methyl-5-isoxazolyl)-3xe2x80x2,4xe2x80x2-methylenedioxyphenyl-4-biphenylsulfonamide and (4-bromo-3-methyl-5-isoxazolyl)-3xe2x80x2-methylphenyl-4-biphenylsulfonamide. Corresponding 4-chloro and 4-fluoro isoxazolyl compounds are also encompassed herein.
Exemplary biphenyl compounds were tested using the exemplified assays (see, EXAMPLES) and the results, which are intended to be exemplary and not limiting, are as set forth in the following table (Table 2):
The preparation of the above and other compounds that possess the requisite activities are set forth in the Examples.
2. Ar2 is Fused Ring Carbocycle or Heterocycle, Including Dibenzofuryl, Dibenzothiophenyl, Dibenzopyrrolyl and Phenanthrene
Compound is in Ar2is a carbocycle containing two or more fused rings or a heterocycle with one heteroatom and two or more fused rings are provided. The heteroatom is O, S or N (NR14) and Ar2 is selected from among, but not limited to, quinolyl, isoquinolyl, dibenzofuryl, dibenzothiophenyl, and dibenzopyrrolyl compounds and other such groups. The fused rings may be substituted with one or more substituents selected from among substituents set forth for R3-R7 or Z above, at any position. The sulfonamide portion of the compounds may be linked at any position on the rings.
In certain embodiments Ar2 is dibenzofuryl: dibenzothiophenyl, dibenzopyrrolyl or phenanthrene or other tricyclic fused ring, and has the following formula: 
in which: R1 and R2 are selected as described above for the thiophenyl, furyl and pyrrolyl compounds; X is xe2x80x94CHxe2x95x90CHxe2x80x94, O, S, NR14, in which R14 is selected independently from the groups set forth above for R11 (compounds in which X is xe2x80x94CHxe2x95x90CHxe2x80x94 are phenanthrenesulfonamides); and
Ar2 is unsubstituted or substituted with one or more substituents selected from R13 and R26, which are each independently (i) or (ii):
(i) R26 and R13 are independently selected from H, OH, OHNH, NH2, NO2, halide, pseudohalide, alkyl, alkenyl, alkynyl, aryl, arylalkyl, heteroaryl, alkoxy, alkylamino, dialkylamino, alkylthio, haloalkoxy, haloalkyl, alkylsulfinyl, alkylsulfonyl, aryloxy, arylamino, arylthio, arylsulfinyl, arylsulfonyl, haloalkyl, haloaryl, alkoxycarbonyl, carbonyl, alkylcarbonyl, aminocarbonyl, arylcarbonyl, formyl, substituted or unsubstituted amido, substituted or unsubstituted ureido, in which the alkyl, alkenyl and alkynyl portions contain from 1 up to about 14 carbon atoms, preferably from 1 to 6 atoms, and are either straight or branched chains or cyclic, and the aryl portions contain from about 4 to about 16 carbons, preferably 4 to 10 carbons; or
(ii) R26 and R13 together are xe2x80x94CH2xe2x80x94, xe2x80x94CHxe2x95x90CHxe2x80x94 O, S, NR11 in which R11 is as defined above, and is preferably, H or alkyl, particularly lower alkyl. It is understood that in either (i) or (ii) each ring of Ar2 may be un-substituted or substituted with more than one substituent, each of which is selected independently from the selections set forth in (i) for R26 and R13.
These compounds, thus, include fused tricyclic-substituted sulfonamides, dibenzothiophenesulfonamides, dibenzofuransulfonamides, dibenzopyrrolesulfonamides (carbazolesulfonamides) and phenanthrenesulfonamides.
In more preferred embodiments, R1 is halide or methyl; R2 is selected from lower alkyl, lower alkenyl, lower alkynyl and lower haloalkyl; R26 and R13 are independently selected from H, lower alkyl, haloalkyl and halide. In more preferred embodiments R1 is Cl, Br or CH3; R2 is selected from H, CH3, C2H5, CF3, n-C3H7, cyclo-C3H7 and C4H9; and R26 and R13 are each independently selected from H, halide, NH2, CF3, CH3, CN, CH3, (CH3)3C, C5H11, CH3O, n-C4H9O and CH2xe2x95x90CH. In yet more preferred embodiments, R2 is H, CH3, C2H5, or CF3; R26 and R13 are independently selected from H, CH3, C2H5, CF3, and halide; and X is O.
Exemplary compounds include those set forth in Table 3:
Preparation of the Compounds
The preparation of the above compounds is described in detail in the examples. Any such compound or similar compound may be synthesized according to a method discussed in general below and by only minor modification of the methods set forth in the Examples by selecting appropriate starting materials as exemplified.
In general, most of the syntheses involve the condensation of a sulfonyl chloride with an aminoisoxazole in dry pyridine or in tetrahydrofuran (THF) and sodium hydride. The sulfonylchlorides and aminoisoxazoles either can be obtained commercially or synthesized according to methods described in the Examples or using other methods available to those of skill in this art (see, e.g., U.S. Pat. Nos. 4,659,369, 4,861,366 and 4,753,672).
The compounds may be prepared by reacting an appropriate sulfonyl chloride with 5-aminoisoxazoles substituted at the 3 and 4 positions (or 4 and 5 positions in the case of 3-isoxazolyl derivatives), such as 5-amino-4-bromo-3-methylisoxazole, in tetrahydrofuran (THF) solution containing a base, such as sodium hydride. Following the reaction, the THF is removed under reduced pressure, the residue dissolved in water, acidified and extracted with methylene chloride. The organic layer is washed and then dried over anhydrous magnesium sulfate, the solvents are evaporated and the residue is purified by recrystallization using hexanes/ethyl acetate to yield pure product. The N-(4-haloisoxazolyl)sulfonamides can be prepared by condensation of amino-4-haloisoxazole with a sulfonyl chloride in THF with sodium hydride as a base.
In some cases, the bis-sulfonyl compound is obtained as the major or exclusive product. The bis-sulfonated products can be readily hydrolyzed to the sulfonamide using aqueous sodium hydroxide and a suitable co-solvent, such as methanol or tetrahydrofuran, generally at room temperature.
Alternatively, the sulfonamides can be prepared from the corresponding sulfonyl chloride and the aminoisoxazole in pyridine with or without a catalytic amount of 4-dimethylaminopyridine (DMAP). In some cases, the bis-sulfonyl compound is obtained as the major or exclusive product. The bis-sulfonated products can be readily hydrolyzed to the sulfonamide using aqueous sodium hydroxide and a suitable co-solvent, such as methanol or tetrahydrofuran, generally at room temperature.
The N-(alkylisoxazolyl)sulfonamides can be prepared by condensing an aminoisoxazole with a sulfonyl chloride in dry pyridine with or without the catalyst 4-(dimethylamino)pyridine. The N-(3,4-dimethyl-5-isoxazolyl)sulfonamides and N-(4,5-dimethyl-5-isoxazolyl)sulfonamides can be prepared from the corresponding aminodimethylisoxazole, such as 5-amino-3,4-dimethylisoxazole. The N-(3,4-dimethyl-5-isoxazolylsulfonamides and the N-(4,5-dimethyl-3-isoxazolylsulfonamdies can be prepared from the corresponding aminodimethylisoxaole, such as 5-amino-3,4-dimethylisoxazole.
Exemplary preparations of numerous compounds herein are set forth in the Examples. For example compounds, such as N-(3,4-dimethyl-5-isoxazolyl)biphenylsulfonamide (EXAMPLE 1), can be prepared from 4-biphenylsulfonyl chloride and an amino-substituted isoxazole, such as 5-amino-3,4-dimethylisoxazole, in dry pyridine (2.0 ml). Following the reaction, the pyridine is removed under reduced pressure and the residue is partitioned between water and ethyl acetate. The organic layer is washed and then dried over anhydrous magnesium sulfate, the solvents are evaporated and the residue is purified by column chromatography over silica gel (e.g., 1% methanol in chloroform as eluent) to yielded a solid. Further purification is achieved by recrystallization from ethyl acetate/hexanes, to yield the pure product. Alternatively, the sulfonamides can be prepared from the corresponding aminoisoxazole in tetrahydrofuran solution containing sodium hydride.
The compounds, such as N-(3,4-dimethyl-5-isoxazolyl)-2-dibenzofuransulfonamide (see, e.g., EXAMPLE 3) can be prepared by reacting 5-amino-3,4-dimethylisoxazole and 2-dibenzofuransulfonyl chloride in dry pyridine. Following the reaction, the pyridine is removed under reduced pressure and the residue is partitioned between water and ethyl acetate. The organic layer is washed and then dried over anhydrous magnesium sulfate, the solvents are evaporated and the residue is purified by column chromatography over silica gel (e.g., 1% methanol in chloroform as eluent) to yield a solid. Further purification is achieved by recrystallization from ethyl acetate/hexanes or column chromatography, to yield the pure product.
Prodrugs and other derivatives of the compounds suitable for administration to humans may also be designed and prepared by methods known to those of skill in the art (see, e.g., Nogrady (1985) Medicinal Chemistry A Biochemical Approach, Oxford University Press, New York, pages 388-392.
Compounds listed and described have been synthesized and tested for activity in in vitro assays and, in some cases, in vivo animal models. Nuclear magnetic resonance spectroscopic (NMR), mass spectrometric, infrared spectroscopic and high performance liquid chromatographic analyses indicated that the synthesized compounds have structures consistent with those expected for such compounds and are generally at least about 98% pure. All of the compounds exemplified or described herein exhibited activity as endothelin antagonists.
C. Evaluation of the Bioactivity of the Compounds
Standard physiological, pharmacological and biochemical procedures are available for testing the compounds to identify those that possess any biological activities of an endothelin peptide or the ability to interfere with or inhibit endothelin peptides. Compounds that exhibit in vitro activities, such as the ability to bind to endothelin receptors or to compete with one or more of the endothelin peptides for binding to endothelin receptors can be used in the methods for isolation of endothelin receptors and the methods for distinguishing the specificities of endothelin receptors, and are candidates for use in the methods of treating endothelin-mediated disorders.
Thus, other preferred compounds of formulas I and II, in addition to those of specifically identified herein, that are endothelin antagonists or agonists may be identified using such screening assays.
1. Identifying Compounds that Modulate the Activity of an Endothelin Peptide
The compounds are tested for the ability to modulate the activity of endothelin-1. Numerous assays are known to those of skill in the art for evaluating the ability of compounds to modulate the activity of endothelin (see, e.g., U.S. Pat. No. 5,114,918 to Ishikawa et al.; EP A 1 0 436 189 to BANYU PHARMACEUTICAL CO., LTD. (Oct. 7, 1991); Borges et al. (1989) Eur. J. Pharm. 165: 223-230; Filep et al. (1991) Biochem. Biophys. Res. Commun. 177: 171-176). In vitro studies may be corroborated with in vivo studies (see, e., U.S. Pat. No. 5,114,918 to Ishikawa et al.; EP A 1 0 436 189 to BANYU PHARMACEUTICAL CO., LTD. (Oct. 7, 1991)) and pharmaceutical activity thereby evaluated. Such assays are described in the Examples herein and include the ability to compete for binding to ETA and ETB receptors present on membranes isolated from cell lines that have been genetically engineered to express either ETA or ETB receptors on their cell surfaces.
The properties of a potential antagonist may be assessed as a function of its ability to inhibit an endothelin induced activity in vitro using a particular tissue, such as rat portal vein and aorta as well as rat uterus, trachea and vas deferens (see e.g., Borges, R., Von Grafenstein, H. and Knight, D. E., Tissue selectivity of endothelin, Eur. J. Pharmacol 165:223-230, (1989)). The ability to act as an endothelin antagonist in vivo can be tested in hypertensive rats, ddy mice or other recognized animal models (see, Kaltenbronn et al. (1990) J. Med. Chem. 33:838-845, see, also, U.S. Pat. No. 5,114,918 to Ishikawa et al.; and EP A 1 0 436 189 to BANYU PHARMACEUTICAL CO., LTD (Oct. 7, 1991); see, also Bolger et al. (1983) J. Pharmacol. Exp. Ther. 225291-309). Using the results of such animal studies, pharmaceutical effectiveness may be evaluated and pharmaceutically effective dosages determined. A potential agonist may also be evaluated using In vitro and in vivo assays known to those of skill in the art.
Endothelin activity can be identified by the ability of a test compound to stimulate constriction of isolated rat thoracic aorta (Borges et al. (1 989) xe2x80x9cTissue selectivity of endothelinxe2x80x9d Eur. J. Pharmacol. 165: 223-230). To perform the assay, the endothelium is abraded and ring segments mounted under tension in a tissue bath and treated with endothelin in the presence of the test compound. Changes in endothelin induced tension are recorded. Dose response curves may be generated and used to provide information regarding the relative inhibitory potency of the test compound. Other tissues, including heart, skeletal muscle, kidney, uterus, trachea and vas deferens, may be used for evaluating the effects of a particular test compound on tissue contraction.
Endothelin isotype specific antagonists may be identified by the ability of a test compound to interfere with endothelin binding to different tissues or cells expressing different endothelin-receptor subtypes, or to interfere with the biological effects of endothelin or an endothelin isotype (Takayanagi et al. (1991) Reg. Pep. 32: 23-37, Panek et al. (1992) Biochem. Biophys. Res. Commun. 183: 566-571). For example, ETB receptors are expressed in vascular endothelial cells, possibly mediating the release of prostacyclin and endothelium-derived relaxing factor (De Nucci et al. (1988) Proc. Natl. Acad. Sci. USA 85:9797). ETA receptors are not detected in cultured endothelial cells, which express ETB receptors.
The binding of compounds or inhibition of binding of endothelin to ETB receptors can be assessed by measuring the inhibition of endothelin-1-mediated release of prostacyclin, as measured by its major stable metabolite, 6-keto PGF1a, from cultured bovine aortic endothelial cells (see, e.g, Filep et al. (1991) Biochem. and Biophys Res. Commun. 177: 171-176). Thus, the relative affinity of the compounds for different endothelin receptors may be evaluated by determining the inhibitory dose response curves using tissues that differ in receptor subtype.
Using such assays, the relative affinities of the compounds for ETA receptors and ETB receptors have been and can be assessed. Those that possess the desired properties, such as specific inhibition of binding of endothelin-1, are selected. The selected compounds that exhibit desirable activities may be therapeutically useful and are tested for such uses using the above-described assays from which in vivo effectiveness may be evaluated (see, e.g., U.S. Pat. Nos. 5,248,807; 5,240,910; 5,198,548; 5,187,195; 5,082,838; 5,230,999; published Canadian Application Nos. 2,067,288 and 2071193; published Great Britain Application No. 2,259,450; Published International PCT Application No. WO 93/08799; Benigi et al. (1993) Kidney International 44:440-444; and Nirei et al. (1993) Life Sciences 52:1869-1874). Compounds that exhibit in vitro activities that correlate with in vivo effectiveness will then be formulated in suitable pharmaceutical compositions and used as therapeutics.
The compounds also may be used in methods for identifying and isolating endothelin-specific receptors and aiding in the design of compounds that are more potent endothelin antagonists or agonists or that are more specific for a particular endothelin receptor.
2. Isolation of Endothelin Receptors
A method for identifying endothelin receptors is provided. In practicing this method, one or more of the compounds is linked to a support and used in methods of affinity purification of receptors. By selecting compounds with particular specificities, distinct subclasses of ET receptors may be identified.
One or more of the compounds may be linked to an appropriate resin, such as Affi-gel, covalently or by other linkage, by methods known to those of skill in the art for linking endothelin to such resins (see, Schvartz et al. (1990) Endocrinology 126: 3218-3222). The linked compounds can be those that are specific for ETA or ETB receptors or other subclass of receptors.
The resin is pre-equilibrated with a suitable buffer generally at a physiological pH (7 to 8). A composition containing solubilized receptors from a selected tissue are mixed with the resin to which the compound is linked and the receptors are selectively eluted. The receptors can be identified by testing them for binding to an endothelin isopeptide or analog or by other methods by which proteins are identified and characterized. Preparation of the receptors, the resin and the elution method may be performed by modification of standard protocols known to those of skill in the art (see, e.g., Schvartz et al. (1990) Endocrinology 126: 3218-3222).
Other methods for distinguishing receptor type based on differential affinity to any of the compounds herein are provided. Any of the assays described herein for measuring the affinity of selected compounds for endothelin receptors may also be used to distinguish receptors subtypes based on affinity for particular compounds provided herein. In particular, an unknown receptor may be identified as an ETA or ETB receptor by measuring the binding affinity of the unknown receptor for a compound provided herein that has a known affinity for one receptor over the other. Such preferential interaction is useful for determining the particular disease that may be treated with a compound prepared as described herein. For example, compounds with high affinity for ETA receptors and little or no affinity for ETB receptors are candidates for use as hypertensive agents; whereas, compounds that preferentially interact with ET_receptors are candidates for use as anti-asthma agents.
D. Formulation and Administration of the Compositions
Effective concentrations of one or more of the sulfonamide compounds of formula I or II or pharmaceutically acceptable salts, esters or other derivatives thereof are mixed with a suitable pharmaceutical carrier or vehicle.
In instances in which the compounds exhibit insufficient solubility, methods for solubilizing compounds may be used. Such methods are known to those of skill in this art, and include, but are not limited to, using cosolvents, such as dimethylsulfoxide (DMSO), using surfactants, such as tween, or dissolution in aqueous sodium bicarbonate. Derivatives of the compounds, such as salts of the compounds or prodrugs of the compounds may also be used in formulating effective pharmaceutical compositions.
The concentrations or the compounds are effective for delivery of an amount, upon administration, that ameliorates the symptoms of the endothelin-mediated disease. Typically, the compositions are formulated for single dosage administration.
Upon mixing or addition of the sulfonamide compound(s), the resulting mixture may be a solution, suspension, emulsion or the like. The form of the resulting mixture depends upon a number of factors, including the intended mode of administration and the solubility of the compound in the selected carrier or vehicle. The effective concentration is sufficient for ameliorating the symptoms of the disease, disorder or condition treated and may be empirically determined.
Pharmaceutical carriers or vehicles suitable for administration of the compounds provided herein include any such carriers known to those skilled in the art to be suitable for the particular mode of administration. In addition, the compounds may be formulated as the sole pharmaceutically active ingredient in the composition or may be combined with other active ingredients.
The active compounds can be administered by any appropriate route, for example, orally, parenterally, intravenously, intradermally, subcutaneously, or topically, in liquid, semi-liquid or solid form and are formulated in a manner suitable for each route of administration. Preferred modes of administration include oral and parenteral modes of administration.
The active compound is included in the pharmaceutically acceptable carrier in an amount sufficient to exert a therapeutically useful effect in the absence of undesirable side effects on the patient treated. The therapeutically effective concentration may be determined empirically by testing the compounds in known in vitro and in vivo systems (see, e.g., U.S. Pat. No. 5,114,918 to Ishikawa et al.; EP A 1 0 436 189 to BANYU PHARMACEUTICAL CO., LTD (Oct. 7, 1991); Borges et al. (1989) Eur. J. Pharm. 165: 223-230;: Filep et al. (1991) Biochem. Biophys. Res. Commun. 177: 171-176) and then extrapolated therefrom for dosages for humans.
The concentration of active compound in the drug composition will depend on absorption, inactivation and excretion rates of the active compound, the dosage schedule, and amount administered as well as other factors known to those of skill in the art. For example, the amount that is delivered is sufficient to treat the symptoms of hypertension. The effective amounts for treating endothelin-mediated disorders are expected to be higher than the amount of the sulfonamide compound that would be administered for treating bacterial infections.
Typically a therapeutically effective dosage should produce a serum concentration of active ingredient of from about 0.1 ng/ml to about 50-100 xcexcg/ml. The pharmaceutical compositions typically should provide a dosage of from about 0.01 mg to about 2000 mg of compound per kilogram of body weight per day. The active ingredient may be administered at once, or may be divided into a number of smaller doses to be administered at intervals of time. It is understood that the precise dosage and duration of treatment is a function of the disease being treated and may be determined empirically using known testing protocols or by extrapolation from in vivo or in vitro test data. It is to be noted that concentrations and dosage values may also vary with the severity of the condition to be alleviated. It is to be further understood that for any particular subject, specific dosage regimens should be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the compositions, and that the concentration ranges set forth herein are exemplary only and are not intended to limit the scope or practice of the claimed compositions.
If oral administration is desired, the compound should be provided in a composition that protects it from the acidic environment of the stomach. For example, the composition can be formulated in an enteric coating that maintains its integrity in the stomach and releases the active compound in the intestine. The composition may also be formulated in combination with an antacid or other such ingredient.
Oral compositions will generally include an inert diluent or an edible carrier and may be compressed into tablets or enclosed in gelatin capsules. For the purpose of oral therapeutic administration, the active compound or compounds can be incorporated with excipients and used in the form of tablets, capsules or troches. Pharmaceutically compatible binding agents and adjuvant materials can be included as part of the composition.
The tablets, pills, capsules, troches and the like can contain any of the following ingredients, or compounds of a similar nature: a binder, such as microcrystalline cellulose, gum tragacanth and gelatin; an excipient such as starch and lactose, a disintegrating agent such as, but not limited to, alginic acid and corn starch; a lubricant such as, but not limited to, magnesium stearate; a glidant, such as, but not limited to, colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; and a flavoring agent such as peppermint, methyl salicylate, and fruit flavoring.
When the dosage unit form is a capsule, it can contain, in addition to material of the above type, a liquid carrier such as a fatty oil. In addition, dosage unit forms can contain various other materials which modify the physical form of the dosage unit, for example, coatings of sugar and other enteric agents. The compounds can also be administered as a component of an elixir, suspension, syrup, wafer, chewing gum or the like. A syrup may contain, in addition to the active compounds, sucrose as a sweetening agent and certain preservatives, dyes and colorings and flavors.
The active materials can also be mixed with other active materials which do not impair the desired action, or with materials that supplement the desired action, such as antacids, H2 blockers, and diuretics. For example, if the compound is used for treating asthma or hypertension, it may be used with other bronchodilators and antihypertensive agents, respectively.
Solutions or suspensions used for parenteral, intradermal, subcutaneous, or topical application can include any of the following components: a sterile diluent, such as water for injection, saline solution, fixed oil, polyethylene glycol, glycerine, propylene glycol or other synthetic solvent; antimicrobial agents, such as benzyl alcohol and methyl parabens; antioxidants, such as ascorbic acid and sodium bisulfite; chelating agents, such as ethylenediaminetetraacetic acid (EDTA); buffers, such as acetates, citrates and phosphates; and agents for the adjustment of tonicity such as sodium chloride or dextrose. Parental preparations can be enclosed in ampules, disposable syringes or multiple dose vials made of glass, plastic or other suitable material.
If administered intravenously, suitable carriers include physiological saline or phosphate buffered saline (PBS), and solutions containing thickening and solubilizing agents, such as glucose, polyethylene glycol, and polypropylene glycol and mixtures thereof. Liposomal suspensions, including tissue-targeted liposomes, may also be suitable as pharmaceutically acceptable carriers. These may be prepared according to methods known to those skilled in the art. For example, liposome formulations may be prepared as described in U.S. Pat. No. 4,522,811.
The active compounds may be prepared with carriers that protect the compound against rapid elimination from the body, such as time release formulations or coatings. Such carriers include controlled release formulations, such as, but not limited to, implants and microencapsulated delivery systems, and biodegradable, biocompatible polymers, such as collagen, ethylene vinyl acetate, polyanhydrides, polyglycolic acid, polyorthoesters, polylactic acid and others. Methods for preparation of such formulations are known to those skilled in the art.
The compounds may be formulated for local or topical application, such as for topical application to the skin and mucous membranes, such as in the eye, in the form of gels, creams, and lotions and for application to the eye or for intracisternal or intraspinal application. Such solutions, particularly those intended for ophthalmic use, may be formulated as 0.01%-10% isotonic solutions, pH about 5-7, with appropriate salts. The compounds may be formulated as aerosols for topical application, such as by inhalation (see, e.g., U.S. Pat. Nos. 4,044,126, 4,414,209, and 4,364,923, which describe aerosols for delivery of a steroid useful for treatment inflammatory diseases, particularly asthma).
Finally, the compounds may be packaged as articles of manufacture containing packaging material, a compound provided herein, which is effective for antagonizing the effects of endothelin, ameliorating the symptoms of an endothelin-mediated disorder, or inhibiting binding of an endothelin peptide to an ET receptor with an IC50 of less than about 10 xcexcM, within the packaging material, and a label that indicates that the compound or salt thereof is used for antagonizing the effects of endothelin, treating endothelin-mediated disorders or inhibiting the binding of an endothelin peptide to an ET receptor.
The following examples are included for illustrative purposes only and are not intended to limit the scope of the invention.